New II. REVIEW OF LITERATUREshodhganga.inflibnet.ac.in/bitstream/10603/6964/6/06... · 2015. 12. 4. · ranged from 10 to 300ppb. High incidence was reported in corn byproducts (57%)

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II. REVIEW OF LITERATURE

Ochratoxin and citrinin are the important fungal toxins that contaminate feeds and feedstuffs and

produces disease condition in domestic animals including poultry. In the present work comparative

pathology of these two toxins have been studied in broiler chicken. The available literature pertinent to the

subject of study is reviewed as hereunder.

2.1 Ochratoxicosis

Ochratoxicosis was first described as a toxic metabolite of Aspergillus ochraceus by Van Der

Merwe et al. (1965), but its complete toxic potential was realized only after isolation of OA from the

sorghum seed strain K-804 in south Africa (Scott, 1965).

Shotwell et al. (1969) first reported the natural contamination of ochratoxin A in corn sample.

Subsequently, Ochratoxin A was also isolated from stored grains (Christensen and Kaufmann, 1969)

including cereals and sorghum (Scott, 1965; Scott et al., 1972), peanuts (Doupnik and Peckham, 1970).

Similarly Penicillium viridactum has been isolated from corn (Krogh and Hasselagar, 1968), wheat (Scott

et al., 1972) and barley (Buckle, 1983). It may be noted that A. ochraceus, has been renamed as Aspergillus

alutaceus.

Huff et al., (1974) opined that ochratoxicosis caused great threat to both human and animal health.

Further, Huff et al., (1975) indicated that ochratoxin has structurally related secondary metabolites

designated as ochratoxin A, B and C of which ochratoxin A (OA) is extremely toxic.

Naturally occurring outbreaks of ochratoxicosis in poultry was reported by Hamilton et al. (1982)

and they investigated five independent episodes of ochratoxicosis in turkeys, two episodes in layers and

two episodes in broiler chicken in United States.

Devi and Polasa (1982) screened 50 maize samples from different feed factories in and around

Hyderabad and reported the presence of ochratoxin in six of them.

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Krogh (1987) and Marquardt et al. (1988) reported occurrence of ochratoxin in different grains,

cereals, plant products, animal feeds and meat from several countries.

Gaur et al. (1991) reported presence of ochratoxin in feed samples from different poultry farms in

Punjab. Raina and Singh (1991) analyzed 42 samples of poultry feed among which nine samples were

positive for ochratoxin.

Egmond and Spijers (1994) reported that cereals and animal meat products were more frequently

contaminated with ochratoxin and their levels ranged from 10 to 500 µg / kg in different ingredients.

Chandrasekaran (1996) noticed high concentration of ochratoxin A in sunflower oil cake (37.0%)

and layer mash (5.6%) in Namakkal area of Tamilnadu.

Devegowda et al. (1998) analyzed various feed ingredients for ochratoxin A collected from

different parts of India and reported an overall incidence of 34.5 per cent and the toxin concentration

ranged from 10 to 300ppb. High incidence was reported in corn byproducts (57%) as compared to oilseed

and oilseed meals.

Thirumala Devi et al. (2002) screened 216 ingredients intended for incorporation into chicken

feeds from 1998-2001 and reported that six per cent of the samples were contaminated with ochratoxin.

Ochratoxin A contamination in excess of 10 µg/kg was found in 9 of 29 sorghum, 1 of 27 groundnut, 1 of

14 rice bran, 1 of 10 sunflower and 2 of 8 millet samples. Ochratoxin was not found in maize, soybean and

mixed feeds.

2.2 Citrinin toxicosis

Citrinin was first isolated from P. citrinum (Hetherington and Raistick, 1931). Soon after its

discovery, citrinin was intended to be used as an antibiotic, but was rejected, because of its marked renal

toxicity. Citrinin has been shown to produce severe renal damage in pigs, rats, dogs and birds and was also

responsible for the ‘Yellowed rice syndrome’, an animal mycotoxicosis reported in Japan (Reiss, 1977).

Citrinin was found mostly in soil and organic substrates, thus contaminating the food grains. It often

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coexisted in cereals along with AF. Its occurrence has been reported in cereal grains, such as corn, wheat,

barley, oats, rye and rice and the levels of contamination ranged from 0.001 to 80 ppm (Roberts and Mora,

1978).

2.3 Toxin Production

2.3.1 Ochratoxin

Ochratoxins are the low molecular weight substances and are produced by several species of

fungus. Six species of Aspergillus viz., A.alliaceus, A.melleus, A.stiamus, A. pertakii, A. sclerotiorum and

A. sulphureus (Lai et al., 1968; Hesseltine et al.,1972) and five species of Pencillium viz., P.commune,

p.cyclopium, P.variable, P.purpurescens and P.palitans are also known to produce ochratoxicosis (Van

Walbeck et al., 1969; Scott et al., 1970; Ciegler et al., 1972). Chu (1974) considered A. Ochraceus as the

principal producer of ochratoxin A. Hamilton et al. (1982) and Dwivedi and Burns (1984a) indicated

Aspergillus ochraceus and Penicillium viridicatum as the principal producer of ochratoxin. However,

Abarca et al. 1994 indicated that ochratoxin A is also produced by A. niger.

2.3.2 Citrinin

Citrinin (CTN) is produced by several species of genus penicillium and Aspergillus namely P.

citrinum, P. viridicatum, P. implicturm, P. fellutanum, P. citreo-viride, P. velutinum, P. purperescens, P.

fenseni, P. spirulosum, P. notatum, P. expansum, P. clasiforms, P. palitans, P. stecki, P. claviforme, P.

roqueforti, A. niveus, A. terreus and A. flavipes (Allman and Steyn, 1979). Citrinin is regarded as a

nephrotoxic mycotoxin. Chemically citrinin, a yellow crystalline substance, is a bicyclic phenol derivative

[(3R-trans)-4,6-dihydro-8hydroxy-3,4,5-trimethyl-6-oxo-3H-2-enzopyran-7-carboxylic acid], with the

molecular formula of C13H14O5 and with a molecular weight of 250. It forms lemon yellow needles (Ciegler

1977). The LD50 value in broiler chicken is 95 mg/kg body weight (Mehdi et al., 1981).

Nelson et al. (1985) reported that nearly 310-632 ppm CTN was extracted from Penicillium spp.

Cultured in corn after various heat treatments. The CTN production in maize was 1050 ppm and 400 ppm

in rice at 25 and 20 per cent of moisture, respectively (Ahamad and Vairamuthu, 2000a).

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2.4 Clinical signs

2.4.1 Clinical signs in ochratoxicosis

Huff et al. (1974) reported clinical signs of listlessness, huddling, diarrhoea, tremors and

prostration within 22 to 25 hours after administration of single oral dose of 16 mg OA per kg feed. Huff et

al. (1975) reported poor growth, increased feed consumption and increased moisture content in manure as

the major clinical signs under field conditions of ochratoxicosis in broilers.

Galtier et al. (1976) observed huddling, hypothermia, cachexia, tremors, ptosis, diarrhoea and

reduction in spontaneous activity during acute short-term toxicity trials with OA in broilers. Decreased feed

consumption, anemia (Huff et al. 1979) and decreased bone strength (Huff and Doerr, 1981) were also

reported in ochratoxicosis of broilers.

Page et al. (1980) observed diarrhoea, paleness of the comb and wattles, emaciation and reduced

water consumption in birds fed with 1.0 ppm of ochratoxin. Where as Hamilton et al. (1982) observed

decreased feed consumption, secondary air sacculitis and mortality both in experimental and natural cases

of ochratoxicosis.

Dwivedi and Burns (1984) reported depressed growth, dullness, huddling, decreased appetite,

reduced feed intake, weakness and diarrhoea in broilers fed with 2 and 4 ppm of OA.

Manning and Wyatt (1984) observed clinical signs of listlessness, emaciation and increased

mortality in chicken fed with diet containing 3 ppm OA. Where as Raina and Singh (1991) reported general

symptoms of dullness, inappetance, reduced feed intake and at times anaemia in broiler chicks fed with diet

containing 1.0 ppm of ochratoxin.

Anorexia, unthriftiness, droopy wings, ruffled feathers, chilled look, neck tucked into the thoracic

cavity and whitish diarrhoea were the predominant signs observed in experimental ochratoxicosis in

broilers (Thyagarajan et al., 1996).

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El-Bagoury and Abdel-Khalek, (1997) observed clinical signs of weakness, closed eyes, loss of

appetite, ataxia and high mortality in a field outbreak of ochratoxicosis in broilers.

Prakash (2001), Rajeev (2001) and Amita (2001) recorded clinical signs of dullness, ruffled

feather, huddling inappetance, diarrhoea, polydypsia, reduced feed conversion and stunted growth in

experimental ochratoxicosis of broilers.

Anil Kumar (2002) and Bhanuprakash (2002) recorded dullness, depression, ruffled feathers

listlessness, huddling, reduced feed consumption, stunted growth and white pasty diarrhoea in birds fed

with 1.0 ppm ochratoxin for five weeks. Similar observations were also made by Santhosh Kumar (2003),

Shivashankar (2004), Anitha (2007) and Ravikumar (2007) in broilers fed with ochratoxin ranging from 1-

2 ppm in the diet.

2.4.2 Clinical signs in citrinin toxicosis

Watery droppings, increased water consumption and decrease in feed intake were the most

consistent clinical sings of citrinin toxicosis and the severity of clinical signs appeared to be dose

dependent when citrinin was fed from 150 to 400 ppm (Tuten 1975; Ames et al., 1976 and Nelson et al.,

1980). Gustavson et al., (1981) reported that citrinin toxicity produced increased water consumption and

urinary excretion. They also indicated that polydipsia could result directly from dehydration and was

associated with polyuria in broiler chicken fed with 200, 400 and 600 ppm citrinin for 12 days.

Mehdi et al. (1981) reported a marked depression, increased water consumption and severe

diarrhoea in broiler chicken administered with 90, 100 and 110 mg of citrinin orally/kg body weight.

Burditt et al. (1983) indicated that P. citrinum contained an unknown factor which caused refusal of feed in

one week-old chicks and citrinin stimulated water consumption. Further, they opined that citrinin caused a

significant growth depression and diarrhoea in broiler chicks fed 130 and 260 ppm up to one week of age.

Uma and Reddy (1995) fed 125 and 250 ppm CTN to broiler chicks from 3 to 5 weeks of age.

Ahamad (1999) fed 150 and 300 ppm citrinin to broiler chicks from 3 to 30 days of age and they reported

loose droppings from first week onwards, which progressed in severity to watery droppings during second

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to fourth week of age with the cumulative exposure to toxin, he noticed progressive anorexia with increased

water intake. Further, during third and fourth week of age, he also observed ruffled feathers, lethargy,

emaciation, marked depression, reduced appetite and stunted growth. Anorexia, poor growth, marked

polydipsia and watery diarrhoea were reported when 150 ppm citrinin was fed to broiler chicks from 1 to

28 days of age (Swaminathan, 2002).

Anandkumar (2006) observed inappetance and brownish diarrhoea from first week onwards and ruffled feathers, lethargy,

and stunted growth from third week in birds fed with 5 ppm citrinin for six weeks.

2.4.3 Clinical signs in combined ochratoxicosis and citrinin toxicosis

Perusal of available literature revealed scanty information on the effect of combined toxicosis on

clinical signs. However, the pertinent literature of combined toxicosis on clinical signs have been reviewed

as hereunder.

Prakash et al. (2002) reported clinical signs of ruffled feathers, listlessness, huddling, inappetance,

poor growth, weight loss, polydypsia and diarrhoea in birds which were fed with 2.0 ppm OA and 1.0 ppm

AF from day-one to thirty five days of age. The authors opined that the intensity of clinical signs was

severe in combined toxicosis when compared to their individual toxicosis. Similar findings were also

observed by Anil Kumar (2002) and Bhanuprakash (2002) in broiler chicks at 500 and 1000 ppb of AF and

OA in the diet, respectively.

Anandkumar (2006) observed clinical sings of inappetance, brownish diarrhoea, reduced appetite, ruffled feathers,

lethargy, and stunted growth in broiler chicken which were fed with 5 ppm CTN and 0.5 ppm AF from day-one to forty two days of

age.

2.5 Mortality

2.5.1 Ochratoxicosis

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Cumulative mortality of 0, 13, 17 and 10 per cent were observed in broiler chicken fed with diets

containing A, Ochraceus contaminated wheat (3mg OA/g diet) and 3.0 µg/g of purified OA in the

potassium salt, sodium salt or organic acid upto four weeks age (Manning and Wyatt, 1984).

Mortality was highest among broilers fed with four mg of OA for three weeks (Gibson et al.,

1989) and nine per cent mortality was observed by Kubena et al. (1994) in male broilers fed with 2 mg

OA/kg of diet upto 19 days of age.

Muthuswamy et al. (2004) reported mortality up to four per cent in broiler chicken fed with 1.0

ppm of OA in the diet. Mortality upto 17 percent was recorded in broilers fed experimental diet containing

ochratoxin level of 4.0 and 2.0 ppm. (Gupta et al. 2005).

Ravikumar (2007) recorded 4 percent mortality in broilers fed experimental diet

containing 1.0 ppm of Ochratoxin.

2.5.2 Citrinin toxicosis

Mehdi et al. (1981) reported 60 per cent mortality in broiler chicken fed with 110 mg CTN/kg

body weight. However, Ahamad (1999) observed no mortality in broiler chicks fed with 150 and 300 ppm

CTN from 3 to 30 days of age. Similarly, Anandkumar (2006) observed no mortality in birds fed with 5.0

ppm citrinin for six weeks.

2.6 Body weight gain


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Sub lethal dose of ochratoxin A and ochratoxin B were reported to cause dose dependent growth

depression in broilers (Peckham et al., 1971). Huff et al. (1974) observed impaired growth rate among

broilers fed 2 ppm OA in the diet.

Prior et al. (1980) reported that the body weights of birds were reduced during the first 5 weeks of

the trial that were received 0.5, 1 and 2 ppm of OA and that the effect was more pronounced among the

males.

Huff and Doerr (1981) reported decreased body weight during second and third week of age in

broilers fed with ochratoxin at 2 g/g in the diet. Manning and Wyatt (1984) reported similar decrease in

body weight in broiler upon feeding ochratoxin at 3 g/g of diet.

Dwivedi and Burns (1984b) recorded growth depression in broilers when fed with 4 ppm OA

while Kubena et al. (1984) observed lowered body weight in broilers fed with ochratoxin A at 0.8 and 1

mg/kg feed.

Singh et al. (1990) and Kumar et al. (2003) reported reduced growth rate and poor feed efficiency

due to presence of ochratoxin A (2 ppm level) in the diet. Broilers fed with ochratoxin at the rate of 3

mg/kg body weight resulted in loss of body weight during fourth week of age (Mohiuddin et al., 1992).

A dose related growth depression was reported by Verma et al. (1995) and Johri et al. (1996) in

broilers fed with 1 to 4 ppm ochratoxin till seven weeks of age. Broilers fed with graded levels of

ochratoxin (0.5, 1.0 and 1.5 ppm/g diet) showed poor feed efficiency (Thyagarajan et al., 1996).

Kubena et al. (1997) reported reduction in the body weight gain by feeding 3ppm ochratoxin to

poults till three weeks of age. Kumar et al. (2003) observed decreased body weight gain and lowered feed

intake in broiler chicks fed with 2 ppm of ochratoxin.

Reduction in body weight gain and increase in FCR was observed in broiler chicks fed with 1.0

ppm of ochratoxin in the diet for five weeks (Anil Kumar et al., 2003; Bhanuprakash et al., 2004).

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Santhosh Kumar (2003) and Shivashankar (2004) reported reduction in body weight and increase

in FCR by feeding 2.0 ppm ochratoxin in broilers up to fifth week of age. Similar observations were also

reported in broilers fed with 0.5 ppm of ochratoxin in the diet (Shivappa, 2005).

Gupta et al. (2005) observed the reduction in body weight in broiler chicken when fed with 2.0

ppm of ochratoxin A in the feed.

Politis et al. (2005) reported an increase in FCR in broiler chicks fed with ochratoxin at the rate of

500 g/kg of diet.

Elaroussi et al. (2006) reported significant depression of body weight in broilers fed with

ochratoxin A at levels of 400 and 800 ppb. Further, they also observed reduction in body weight in broiler

chicken when fed with 1.0 ppm of ochratoxin.


Ames et al.(1976) reported that feeding citrinin at 500 mg/kg diet significantly depressed the body

weight gain of broilers by first week of the experiment and the effect continued up to third week. Roberts

and Mora (1979) reported that broiler chicks fed with 130 and 260 ppm CTN up to six weeks of age

showed significant growth depression. Growth depression was also reported in broilers by feeding different

levels of citrinin viz., 500 ppm for 1 to 3 weeks (Campbell et al., 1981), 220, 330 and 440 ppm for three

weeks (Nelson et al., 1981), 300 ppm for 14 and 20 days (Manning et al., 1985) 125 and 250 ppm for 3 to 5

weeks of age (Uma and Reddy, 1995), 150 and 300 ppm levels from 3rd to 30th day (Ahamad, 1999) and at

150 ppm from 1 to 28 days of age (Swaminathan, 2002).

Anandkumar (2006) observed decrease in body weight gain from second week onwards in broiler chicken fed with 5.0

ppm of citrinin for six weeks.

2.6.3 Combined Ochratoxicosis and citrinin toxicosis

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body weight gain. However, the pertinent literature of combined toxicosis on body weight gain has been

reviewed as hereunder.

Ahamad (1999) reported significant reduction in body weight gain in broiler chicks fed with 150

ppm CTN and 0.5 ppm AFB1 from 3 to 30 days of age as compared to the control group.

2.7 Feed consumption and conversion


Hamilton et al. (1977) observed reduced feed consumption among broilers in field cases of

ochratoxicosis.

Prior et al. (1980) recorded reduced feed consumption among birds which received OA at 0.5,

1and 2ppm level during first five weeks of the trial. All metabolites of OA at 3 ppm dose level caused

significant increase in cumulative feed conversion ratio in broilers (Manning and Wyatt., 1984).

Kubena et al. (1988) observed lowered feed efficiency in male broilers fed with 2 ppm OA from 1

to 3 weeks of age. Increased feed conversion ratio was also recorded in OA fed 3 week old broilers

(Gibson et al. 1989).

Niemiec and Scholtyssek (1989) reported poor feed conversion in broilers that received 1.5ppm

OA for six weeks in the diet. Poor feed efficiency was observed in broilers fed with graded levels of OA (1-

4 ppm) till seven weeks of age (Verma et al., 1995).

Verma et al. (1995) observed reduced feed intake when OA (2 and 4 ppm) was fed to broilers.

Gentles et al. (1999) reported decreased feed efficiency in broilers fed with 2.5 ppm OA. A

significant reduction in body weight and poor feed conversion was found when broilers were fed with 2.0

ppm OA in the diet (Raju and Devegowda, 2000).

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Anil Kumar (2002) observed increased feed consumption ratio in birds fed with 1.0 ppm

ochratoxin for 5 weeks. Similarly, Kumar et al. (2003) observed decreased body weight gain and reduced

feed intake in broiler chicks fed with 2 ppm ochratoxin.

Santhosh Kumar (2003) reported reduction in body weight and increase in FCR by feeding 2.0

ppm ochratoxin in broilers up to fifth week of age.

Reduction in body weight gain and increase in FCR was observed in broiler chicks fed with 1.0

ppm ochratoxin in the diet for five weeks (Anil Kumar et al., 2003; Bhanuprakash et al., 2004).

Gupta et al. (2005) reported poor feed conversion in broilers receiving a diet containing 2 and 4

ppm OA for six weeks. Similar observations were also made in broilers with 0.5 ppm of ochratoxin

(Shivappa, 2005).

Anitha (2007) and Ravikumar (2007) observed reduction in feed consumption in broiler chicken fed with 1.0 ppm

ochratoxin for 5 weeks.


Ames et al. (1976) reported decreased feed consumption in broilers fed 62.5, 125, 250 and 500

ppm CTN for three weeks. Nelson et al. (1981) recorded decrease in feed efficiency ratio in male broiler

chicks fed with 440 ppm CTN for three weeks. Doerr et al. (1982) observed reduced feed consumption,

however, feed conversion ratio remained unaffected when broiler chicks were fed with 440 ppm CTN for

three weeks. A relative decrease in feed consumption and increase in water intake with no significant

difference in feed conversion ratio were observed in broiler chicks fed with 125 and 250 ppm CTN up to

six weeks of age (Uma and Reddy, 1995).

Citrinin, when fed to broilers at 150 and 300 ppm levels from 3 to 30 days of age (Ahamad, 1999)

and 150 ppm level from 1 to 28 days of age (Swaminathan, 2002) caused significant decrease in feed

consumption, but not in feed conversion.

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Anandkumar (2006) observed significant reduction in feed consumption in broiler chicken fed

with 5.0 ppm of citrinin in the diet.

2.7.3 Combined ochratoxicosis and citrinin toxicosis


feed consumption and conversion. However, the available pertinent literature of combined toxicosis on feed

consumption and conversion have been reviewed as hereunder.

Ahamad (1999) reported significant decrease in feed consumption, but not in feed conversion

when broiler chicks were fed 150 ppm CTN along with 0.5 ppm AFB1 from 3 to 30 days of age.

2.8 Haematological parameters


Perusal of literature on haematological study in ochratoxicosis did not reveal any pertinent

literature to review.


Uma and Reddy (1995) reported no significant change in packed cell volume (PCV), haemoglobin

(Hb) percentage and total erythroxyte count (TEC) in broiler chicken fed with 125 and 250 ppm CTN up to

six weeks of age. Similarly, no significant difference was observed in PCV, Hb and TEC when CTN was

fed to broiler at 150 and 300 ppm levels, from 3rd to 30th day of age (Ahamad, 1999) and at 150 ppm level

from 1 to 28 days of age (Swaminathan, 2002). However, Anandkumar (2006) observed significant

decrease in PCV, Hb and TEC in broiler chicken fed with 5.0 ppm of citrinin in the diet.

2.8.3 Combined ochratoxicosis and citrinin toxicosis

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haematological parameters. However, the pertinent literature of combined toxicosis on haematological

parameters have been reviewed as hereunder.

Broiler chicks fed 150 ppm CTN and 0.5 ppm AFB1 from 3 to 30 days of age showed highly

significant decrease in PCV, Hb and TEC values when compared to the control and CTN alone treated

groups (Ahamad, 1999).

2.9 Serum biochemistry


2.9.1.1 Serum total protein, albumin and globulin

Manning and Wyatt et al. (1984) reported significantly low levels of total protein, albumin and globulin by feeding three

g of ochratoxin A per gram of diet. Similar observation with two mg of ochratoxin per kg of diet was also reported by Kubena et al.

(1988).

Huff et al. (1988) reported low levels of total protein and albumin in birds upon feeding ochratoxin at dose level of 2 and 4

g/g of feed throughout the experimental period for 21 days.

Singh et al. (1990) observed significant decrease in total protein, albumin and globulin levels at the dose rate of 0.5 and 2.0

ppm ochratoxin A in broilers.

Many earlier studies have also indicated decrease in total protein, albumin and globulin levels (Mohuiddin et al., 1992;

Kubena et al., 1994; Gentles et al., 1999; Prakash et al., 2002; Kumar et al., 2003) during ochratoxicosis in broiler chicken.

Gupta et al. (2005) reported significant reduction in the levels of serum total protein, albumin and globulin in broiler

chicken fed with 1.0 ppm ochratoxin for 6 weeks.

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A significant reduction in serum concentration of total protein, albumin and globulin were recorded in birds that consumed

1 ppm ochratoxin for 5 weeks (Anitha 2007 and Ravikumar 2007).

2.9.1.2 Glucose

Perusal of literature revealed no pertinent reference on the effect of ochratoxin on glucose levels in

birds.

2.9.1.3. Serum alanine transaminase

Raina and Singh (1991) reported increase in the activity of Serum alanine transaminase (ALT) in broiler chicken fed with

1.0 ppm of ochratoxin in the diet.

Verma et al. (1995) observed an increase in the activity of ALT enzyme in broiler chicken fed with 1.0 to 4.0 ppm

ochratoxin in the feed.

Raju and Devegowda (2000) observed increase in the serum ALT values in broilers fed with 2.0 ppm ochratoxin in the

diet. Similarly, elevated serum ALT enzymes with ochratoxin in broilers were reported by Kumar et al. (2003) and Gupta et al.

(2005).

Anitha (2007) and Ravikumar (2007) observed increase in serum ALT values in broilers fed with 1 ppm of ochratoxin in

the diet.

2.9.1.4 Serum aspartate transaminase

Kubena et al. (1988) observed a decrease in Serum aspartate transaminase (AST) level in ochratoxin treated broilers.

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Mohiuddin et al. (1992) reported no change in the serum AST level when ochratoxin was fed at the rate of 0.75, 1.50 and

3.00 g/kg body weight in broiler chicken.

An increase in serum AST level was reported when 2 ppm of ochratoxin was fed to broilers (Kumar et al., 2003 and Gupta

et al., 2005).

Anitha (2007) and Ravikumar (2007) observed an increase in serum AST values in broilers fed with 1 ppm of ochratoxin in

the diet.

2.9.1.5 Serum alkaline phosphatase

Huff et al. (1988) reported no dose related alteration in ALP levels of broilers fed with 2 and 4 ppm of ochratoxin.

Kubena et al. (1988) and Raina and Singh (1991) observed significant increase in serum alkaline phosphatase (ALP) level

in broilers fed with ochratoxin contaminated diet.

Serum activity of enzyme ALP was significantly increased with 2 ppm of ochratoxin in broilers (Gupta et al., 2005).

Anitha (2007) observed increase in serum ALP values in broilers fed with 1 ppm of ochratoxin in the diet.

2.9.1.6 Serum creatinine

Huff et al. (1988) reported significantly increased creatinine level when 2 and 4 g dose level of ochratoxin was fed to

broilers.

Sakthivelan and George (2002) indicated that the creatinine levels were uniformly elevated when ochratoxin was fed at the

dose of 2 mg/kg feed in broilers.

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Elevated creatinine level was observed in broilers fed with 2 ppm ochratoxin in the diet (Kumar et al., 2003 and Gupta et

al., 2005).

Anitha (2007) and Ravikumar (2007) observed increase in serum creatinine values in broilers fed with 1 ppm of ochratoxin

in the diet.

2.9.1.7 Uric acid

Huff et al. (1975) recorded increased levels of uric acid in broilers when they were fed with 4 and

8 ppm OA in the diet.

Mohiuddin et al. (1992) observed a significant increase in uric acid level in birds fed with 3 ppm

OA for 4 weeks.

Elevated serum uric acid levels were reported in broiler chickens administrated with 2.0 ppm

ochratoxin in the feed (Prakash et al., 2002; Rajeev, 2001; Amita, 2001 and Shivashankar, 2004).

Ravikumar (2007) observed increase in serum uric acid values in broilers fed with 1 ppm of ochratoxin in the diet.


Ames et al. (1976) reported that feeding up to 500 ppm CTN for three weeks in broiler chicken

had no effect on serum protein, glucose, cholesterol, uric acid and calcium or potassium levels. In broiler

chicks fed with 400 ppm CTN for three weeks, the total protein and albumin levels were not affected

(Doerr et al., 1982). Manning et al. (1985) reported that feeding broiler chicken with 300 ppm CTN for

three weeks resulted in significant increase in serum total protein, albumin, globulin, glucose and

cholesterol, but no change in the uric acid level.

Uma and Reddy (1995) reported that the serum glucose, total protein and uric acid values were not

significantly altered in broiler chicks fed with 125 and 250 ppm CTN for six weeks. Ahamad (1999)

reported no significant difference in serum total protein, when CTN was given to broiler chicks at 150 and

86

300 ppm levels. In broiler chicks fed with 150 ppm CTN from 0 to 28 days of age a significant increase in

the serum albumin, cholesterol and glucose and a non-significant increase in serum proteins were observed

when compared to control, whereas Swaminathan (2002) reported no change in the serum uric acid,

creatinine and alkaline phosphatase (ALP) levels in CTN fed birds.

Anandkumar (2006) observed decreased serum total protein, albumin, globulin, glucose, calcium,

sodium and increased ALT, BUN, triglyceride level in broiler chicken fed with 5.0 ppm citrinin through

diet.

2.10 Relative organ weight


Huff et al. (1974) reported a significant increase in the relative weight of liver at higher dose of 8

g/g and increase in weight of kidney even at a lower dose of 1 g/g diet. They also observed significant

increase in the weight of proventriculus and gizzard at doses of four and eight g/g, respectively.

Hamilton et al. (1982) and Manning and Wyatt (1984) observed increase in weight of liver and

kidneys in broilers fed with 2.0 ppm of ochratoxin.

Dwivedi and Burns (1984) observed marked enlargement of kidneys to about one and half times

the normal size at 2 ppm and almost twice the size of control when 4 ppm ochratoxin was administered.

Further, they also observed significant decrease in size of thymus and bursa of Fabricius with 2 and 4 ppm

ochratoxin in the diet.

Kubena et al. (1986) reported an increase in the relative weights of liver, kidneys, proventriculus

and gizzard but no difference in the relative weights of the spleen and bursa of Fabricius at a dose level of

2.5ppm ochratoxin in feed. Similar results were also observed by earlier workers in experimental

ochratoxicosis in broilers (Kubena et al., 1989; Huff et al., 1992).

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Several workers (Huff et al., 1992; Verma et al., 1995 and Kubena et al., 1997) have reported an

increase in the relative weights of kidneys, liver and other organs after feeding ochratoxin contaminated

feed.

Ochratoxin A at 2.5 ppm in diet produced an increase in relative weight of kidneys in broilers

(Gentles et al., 1999). Similar response of increase in mean liver, kidney and spleen weight, and reduction

in weight of thymus and bursa of Fabricius were observed when ochratoxin was incorporated in the diet at

2ppm level (Prakash, 2001; Rajeev, 2001).

Amita (2001) found reduction in relative weights of bursa of Fabricius and thymus and increase in

relative weights of liver and kidneys in birds fed with 2ppm of ochratoxin A in diet.

Sakthivelan and George (2002) reported an increase in weight of kidneys and liver and reduction

in the weight of bursa of fabricius, thymus and spleen in chicks fed with 2ppm ochratoxin in the diet.

Stoev et al. (2002) recorded an increase in the weight of liver and kidneys and decrease in the

weight of bursa of Fabricius, thymus and spleen in chicks fed with ochratoxin contaminated diet.

Anilkumar (2002) and Bhanuprakash (2002) reported increase in weight of kidneys and liver and

decrease in the weight of bursa of Fabricius, thymus and spleen in chicks fed with 1.0 - 2.0 ppm levels of

ochratoxin in the diet (Santhosh Kumar, 2003; Shivashankar, 2004).

Elaroussi et al. (2006) reported an increase in the relative weight of gizzard and decrease in

relative weight of thymus in broilers fed with 400 and 800 ppb of ochratoxin.

Anitha (2007) and Ravikumar (2007) observed significant increase in the relative

weights of liver and kidneys and decrease in the relative weights of lymphoid organs in

broiler chicken fed with 1.0 ppm of ochratoxin.


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Ames et al. (1976) reported increase in the relative kidneys and liver weights at the highest dose

level in broiler chicks fed with 62.5, 125, 250 and 500 ppm CTN for three weeks. Kidneys weights were

significantly increased, but bursa of Fabricius, pancreas, liver, spleen and thymus were unaffected in broiler

chicks fed with 125 ppm CTN for three weeks (Campbell et al., 1981).

Broiler chicks fed with 300 µg CTN/g from the day of hatch to three weeks of age showed

increased relative weights of liver and kidneys. The increase in kidney weight of CTN group was to the

tune of 37% (Manning et al. 1985). Liver and kidneys of broiler chicks fed with 150 and 300 ppm CTN

from 3 to 30 days (Ahamad, 1999) and 150 ppm from 11 to 28 days (Swaminathan, 2002) evinced a

significant increase in their weights.

Anandkumar (2006) observed increase in the relative weights of liver and spleen and decrease in

the weight of bursa of Fabricius in broiler chicken fed with 5.0 ppm of citrinin in the diet.

2.11 Immunity

2.11.1 Humoral immunity

2.11.1.1 Ochratoxicosis

Dwivedi and Burns, (1984) documented significant decrease in immunoglobulin containing cells

in all the lymphoid organs studied. Correspondingly, deposition of immunoglobulins, especially IgG in

glomerular basement membrane. The total immunoglobulin levels were also reduced in the sera of birds fed

with 2 and 4 ppm ochratoxin. Reduction in HI titers against Newcastle disease was recorded as the dietary

level of ochratoxin was increased from 0-1.5 ppm in broiler diets (Thyagarajan et al., 1996). Significant

reduction in spleen weight was reported in broilers which received 3 ppm ochratoxin (Kubena et al., 1997).

Stoev et al., (2000) noticed lower haemagglutination inhibition antibody titers and reduced

relative weight of lymphoid organs in broiler chicks fed with ochratoxin ranging from 130 to 790 ppb in the

diet.

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Anil Kumar (2002) reported significant reduction in HI titer against NDV in birds fed with 1 ppm

of ochratoxin in the diet.

2.11.1.2 Citrinin toxicosis

Day-old broiler chicks fed with 125, 250 and 500 ppm CTN for three weeks and immunized at 22

days of age with sheep red blood cells or killed Brucella abortus antigen produced total agglutinating

antibody titres equivalent to control chicks, on seventh day. Mercaptoethanol resistant (IgG) and sensitive

CTN did not interfere with humoral-mediated immune functions (Campbell et al., 1981). In broiler chicks

fed with 400 ppm CTN from day-old to three weeks of age, the globulin levels were depressed (Doerr et

al., 1982).

In broiler chicks fed with 5 ppm CTN from day-one to six weeks of age, the globulin levels were

depressed, the overall mean ND-HI titres were decreased significantly in the CTN group when compared to

the control group (Anandkumar, 2006).

2.11.2 Cell mediated immunity


Chang and Hamilton (1980) reported decreased phagocytic and locomotory capacities of

heterophils in chicks exposed to dietary OA.

Dwivedi and Burns (1984) found a dose-related regression of bursa of Fabricius, thymus and

spleen with ingestion of OA. Cell mediated immune response, as measured by delayed hypersensitivity

skin reaction to avian tuberculin and to bovine serum albumin (BSA) in pre sensitized birds were

significantly depressed in growing turkey fed 4 ppm dietary ochratoxin A.

Singh et al., (1992) found highly significant reduction in CMIR by way of diminished skin

sensitivity, graft versus host reaction, T lymphocyte counts and the number of nitroblue tetrazolium test

positive cells in broilers fed with 0.5 and 2 ppm ochratoxin.

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Significant reduction in mean skin fold thickness at 24 and 48 hours post treatment with DNCB

was observed in birds fed with OA at 1 ppm in the diet when compared to control (Anil Kumar, 2002).


Campbell et al. (1981) fed broiler chicks with 125, 250 and 500 ppm CTN for three weeks and

observed normal differential count and reported that there was no effect of CTN on the capacity of

heterophils to phagocytize bacteria, which suggested that CTN did not interfere with cell mediated

immunity.

2.12 Gross pathology


Peckham et al. (1971) reported extensive accumulation of urates on the serosal surface of visceral

organs in chicks after an oral administration of OA in the range of 0.1 to 100µg/ chick.

Huff et al. (1974) observed enlarged kidneys, proventiculus, gizzard, liver and atrophy of bursa of

Fabricius among broiler chicken fed with 0.5, 1, 2, 4 and 8 µg of OA /g of diet for 3 weeks.

Dwivedi and Burns (1984a) observed pale, enlarged kidney, liver and proventriculus along with

less viscous bile and distended gall bladder in broiler chicks fed with 2 and 4 ppm of ochratoxin

contaminated diets.

Dietary inclusion of ochratoxin at the rate of 3 g/kg body weight resulted in enlarged and

congested liver, distended gall bladder with full of bile and slightly atrophied kidneys (Mohiuddin et al.,

1992).

Emaciation, dehydration, dry and firm gizzard with erosion on the proventricular mucosa,

haemorrhages and catarrhal enteritis were the lesions observed in growing chicks which received OA at the

levels of 2 and 4ppm for 20 days (Leeson et al., 1995).

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Thyagarajan et al. (1996) observed pale kidneys with chalky white urate deposits on the surface of

kidneys, heart, mesentery, liver, peritoneum and pericardium, catarrhal enteritis and intestinal dilatation in

broilers fed with ochratoxin at the rate of 0.5, 1.0 and 1.5 g /g of diet.

Ramadevi et al. (1998) recorded pale, enlarged liver and kidneys with petechial haemorrhages,

catarrhal enteritis, congestion and focal haemorrhages in pancreas and mucus accumulation in

proventricular lumen in broilers fed with 1 and 2 ppm OA for 6 wks. of age. Atrophy of thymus and bursa

of Fabricius with enlargement of spleen was noticed in broilers fed with 2ppm ochratoxin in feed (Prakash,

2001; Rajeev 2001).

Amita (2001) and Sakthivelan and George (2002)observed pale, enlarged liver and kidneys,

distended gall bladder, regressed bursa of Fabricius and thymus in birds fed with 2ppm of ochratoxin.

Elizabeth et al. (2002) noticed pale discoloration of the feet and beak, pale swollen kidneys and

yellowish liver when broilers were administered with 2 ppm of ochratoxin in the diet.

Pale enlarged liver and kidneys; distended gall bladder, regressed bursa of Fabricius and thymus

in broilers that received 1.0 and 2.0 ppm ochratoxin in the feed (Anilkumar 2002, Bhanuprakash 2002,

Santhosh Kumar 2003, Anitha 2007 and Ravikumar 2007).


Ames et al. (1976) observed enlargement of liver and kidneys in broiler chicks fed 62.5 to 500

ppm CTN for three weeks. Roberts and Mora (1978) reported that broiler birds fed with 130 and 260 ppm

CTN evinced haemorrhagic jejunum, mottled liver, enlarged kidneys and distended crop.

Mehdi et al. (1981) reported that birds fed with 90, 100 and 110 ppm CTN by oral gavages

exhibited moderately enlarged and congested kidneys and liver. Pale yellow to gray and swollen kidneys

were observed, when CTN was fed orally at the rate of 50, 70 and 80 mg/kg body weight as single dose in

turkey poults and 30, 40 and 50 mg/kg body weight as single oral dose in ducklings (Mehdi et al., 1983).

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Uma and Reddy (1995) recorded enlargement, congestion and mottling of kidneys, enlargement,

congestion and fragility of liver, petechiae on fascia and thigh muscle and dilatation of heart in broiler

chicken fed with 125 and 250 ppm CTN for six weeks.

Petechiae on kidneys, epicardium, myocardium, fascia and thigh muscles were observed in broiler

chicks fed 300 ppm CTN from 3 to 30 days of age (Ahamad and Vairamuthu, 2001). Swaminathan (2002)

reported enlargement, congestion and in a few birds, pale and pliable liver, distended gall bladder with

greenish, thick bile, mild to moderate enlargement of kidneys, moderate to marked distension of caecum

with pasty and grayish brown coloured feces. In addition, they also recorded hemorrhagic caecal tonsils,

congestion of bursa of Fabricius in broiler chicks fed with 150 ppm citrinin from 1 to 28 days of age.

Anandkumar (2006) observed lesions of congestion, enlargement, paleness or yellowish

discolouration of liver, distended gall bladder; enlargement, congestion and petechiae on kidneys.

Splenomegaly, atrophy of bursa of Fabricius and catarrhal enteritis in broilers fed with 5 ppm citrinin in the

diet.

2.13 Histopathology

2.13.1 Liver


Peckham et al. (1971) observed focal necrosis of hepatocytes in broiler chicken fed with OA at the

dose level of 4 ppm.While Huff et al. (1979) reported increased intracytoplasmic deposits of glycogen

especially in the hepatocytes at the periphery of liver lobes in broiler chicken fed with diet containing 4 and

8 ppm OA. Pepeljnjak et al. (1982) reported adenomatous hyperplasia of hepatic parenchyma in OA

treated birds.

Degeneration and vacuolation of hepatocytes, perivascular inflammatory reaction and nodular lymphoid infiltration along

with congestion and haemorrhages were noticed in chicks fed with 2 and 4 ppm of ochratoxin A for three weeks (Dwivedi and Burns,

93

1984). Mild hepatocellular degeneration with diffuse lipidosis of the liver was observed in ochratoxin treated birds (Kubena et al.,

1988 and Raina and Singh, 1991).

Mohiuddin et al. (1992) recorded congestion of sinusoids and central vein, focal necrosis with

mononuclear cell infiltration and fatty changes in liver when broilers were fed with ochratoxin at the rate of

3 mg /kg diet for 8 wks.

Ramadevi et al. (1998) observed congestion of vessels, fatty changes in hepatocytes, bile duct proliferation, focal lymphoid

aggregation and necrosis of hepatocytes in broilers fed with 1.0 ppm of ochratoxin through diet.

Roy et al. (2000) observed focal necrosis of hepatocytes in natural cases of ochratoxicosis in

grower chickens due to consumption of OA contaminated drinking water.

Prakash (2001) and Rajeev (2001) reported congestion of vessels and sinusoidal spaces, vacuolar

degeneration, infiltration of heterophils and lymphoid cells in the portal triad in ochratoxin (2 ppm) fed

broilers.

Congestion of vessels and sinusoidal spaces, hydropic to fatty changes of hepatocytes, biliary

hyperplasia, heterophic infiltration and focal areas of lymphoid aggregates were the lesions observed in the

liver of broilers fed with 2ppm ochratoxin for four weeks (Amita et al., 2001).

Sakthivelan and George (2002) observed sinusoidal congestion, hepatocyte vacuolation, scattered

necrotic foci, multifocal heterophilic infiltration with fibroplasia, acinar transformation of hepatocytes and

bile duct hyperplasia in broilers fed with ochratoxin at 1 and 2ppm levels.

Krisztina et al. (2002) noticed discrete focal fibrosis around bile duct and mild epithelial proliferation of biliary canaliculi

in broilers exposed to 0.5 mg of ochratoxin A.

Anilkumar (2002), Bhanuprakash (2002), Santhosh Kumar (2003), and Shivashankar (2004)

reported congestion of the vessels and sinusoidal space, mild degree of hydropic and fatty change, biliary

94

hyperplasia and periportal lymphocytic infiltration in broiler chicken fed with 1.0 and 2.0 ppm ochratoxin

in the diet for five weeks.

Aravind Kumar et al. (2004) noticed congestion and haemorrhages degeneration of hepatocytes with fatty changes in

broilers fed with 2 ppm ochratoxin in the diet.

Anitha (2007) observed congestion, haemorrhages, hydropic degeneration and

biliary hyperplasia with periportal infiltration of heterophils in the liver of birds fed with

1.0 ppm of ochratoxin.

Ravikumar (2007) observed vacuolar degeneration of cytoplasm, mild fatty

changes, necrosis of hepatocytes, increased kupffer cell activity, focal areas of

inflammatory cells infiltration and bile duct epithelial hyperplasia in liver of birds fed

with 1.0 ppm of ochratoxin.


Roberts and Mora (1978) observed hydropic degeneration and coagulative necrosis of portal and

centrilobular hepatocytes, dilatation of central vein and hyperplastic bile ducts in broiler chicks fed 130 and

260 ppm CTN for six weeks. Degenerative changes and lymphocytic aggregation in liver were found in all

broiler chicks fed with 33, 65, 135 and 260 ppm CTN for six weeks (Roberts and Mora, 1979).

Mehdi et al. (1981) observed multiple foci of necrosis and haemorrhages in liver of chicks fed

with 47.5 to 500 ppm CTN. The hepatocytes showed pale, frequently vacuolated eosinophilic cytoplasm,

pyknotic, and karyorrhectic or lysed nuclei in chicks fed with 90, 100 and 110 ppm CTN in single oral

dose.

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Maryamma et al. (1990) observed mild focal degenerative lesions which consisted of granular and

fatty changes in cytoplasm of some of the peripheral hepatocytes when fed with 125 mg CTN/kg body

weight by crop intubation in chicken. Uma and Reddy (1995) observed mild to moderate congestion with

dilatation of central vein and sinusoids, fatty changes in the hepatic cells, focal proliferation of Kupffer

cells and mononuclear cell infiltration in the portal triads when chicks were fed with 125 and 250 ppm

CTN for six weeks.

In broiler chicks fed with 150 and 300 ppm CTN from 3 to 30 days of age (Ahamad and

Vairamuthu, 2001), liver evinced mild to moderate sinusoidal congestion, diffuse vacuolar degeneration of

hepatocytes with multifocal mononuclear and heterophilic infiltration. Bile duct hyperplasia was observed

in most of the broiler chicks fed with 150 ppm CTN for 28 days of age (Swaminathan, 2002).

Anandkumar (2006) observed congestion of vessels, ballooning and vacuolar degeneration of

hepatocytes, micro and marcovesicular fatty degeneration and acinar transformation of hepatocytes. He

also observed Kuffer cell hypertrophy in the third week and immense periductular mononuclear cell

infiltration during the sixth week of age in birds fed with 5.0 ppm of citrinin.

2.13.2 Kidneys


Peckham et al. (1971) reported acute nephrosis with gout, presence of proteinaceous cast, urates,

scattered heterophils and localized necrosis in renal tubules of chicks fed with both Ochratoxin A and

Ochratoxin B. Whereas, Huff et al. (1975) observed swelling of tubular epithelial cells, tubular dilatation

with the presence of proteinaceous material in the lumen of proximal convoluted tubules (PCT) in the OA

treated birds at the levels of 4 and 8 µg OA/g feed. Galtier et al. (1976) found microhaemorrhages in both

cortex and medulla of kidney in OA treated broiler chicks.

Page et al. (1980) reported that the lesions were confined to the tubular epithelium of the kidney in ochratoxicosis. The

changes included granular appearance of tubular epithelium along with hyperplasia of tubular cells.

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Dwivedi and Burns (1984) observed that the OA principally affected the PCT of the kidney of

broiler chicken causing severe distension, enlargement and hypertrophy with the presence of hyaline

globules in the lumen. They also reported necrosis of individual cells of PCT, presence of mitotic figures

and the thickening of glomerular basement membrane which was often accompanied by swelling of the

glomerular capillary endothelium.

Mohiuddin et al. (1992) observed congestion of the capillaries, degenerative changes of epithelial

cells of convoluted tubules and the loss of brush border in the collecting tubules of kidneys in broilers fed

with ochratoxin at the rate of 3 mg / kg for eight weeks period.

Tubular degeneration and adenomatous hyperplasia were observed in experimental ochratoxicosis of broilers (Thyagarajan

et al., 1996).

Ramadevi et al. (1998) reported congestion, degeneration, hyperplasia and hypertrophy of the

cells of glomeruli, focal interstitial nephritis, mild fatty changes, intertubular fibroblastic proliferation,

cystic dilatation of the tubules with occasional hyaline casts and periglomerular fibrosis with thickening of

basement membrane in broiler chicken treated with 1 and 2ppm OA from day 1 to 6 weeks of age. While,

Roy et al. (2000) found tubular dilatation, marked infiltration of heterophils, localized necrosis of renal

tubules and atrophy of glomeruli due to consumption of OA contaminated drinking water in grower

chicken.

Prakash (2001) and Rajeev (2001) observed loss of brush border of PCT, misshapened tubules,

vacuolar degeneration, necrosis and presence of fibrinous deposits in the lumen of ochratoxin fed birds.

While Amita (2001) recorded lesions such as congestion haemorrhages, degenerative changes, loss of brush

border of PCT, dilatation and misshapened tubules with presence of eosinophilic hyaline casts in the lumen

of broiler chicken fed with 2 ppm ochratoxin.

Sakthivelan and George (2002) recorded dilatation of tubules with occasional hyaline casts,

intertubular haemorrhages, mononuclear cell infiltration and glomerular haemorrhages in broilers fed with

ochratoxin levels ranging from 1 to 2 ppm in the diet.

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Krisztina et al. (2002) noticed enlarged glomeruli, swollen capillary endothelial cells, thickened glomerular basement

membrane, degenerative tubules and multiple adenoma like lesions in renal parenchyma in broiler chicks exposed to 0.5 mg of

ochratoxin.

Congestion of vessels, multifocal areas of haemorrhages, distended PCT with loss of brush border

and cytoplasmic vacuolations were noticed in broiler chicken fed with 1.0 ppm of ochratoxin in the diet

(Anilkumar 2002 and Bhanuprakash, 2002).

Santhosh Kumar (2003) and Shivashankar (2004) reported loss of brush border of PCT, dilatation

and misshapened tubules, vacuolar degeneration and necrosis, presence of fibrinous deposits in the lumen

of broiler chicken and ducklings fed with 2.0ppm ochratoxin for five weeks.

Anitha (2007) observed distended, misshapen PCT with loss of brush border and presence of proteinaceous material in the

lumen, degenerative changes of glomeruli, stray collection of heterophils and lymphocytes in the interstitium along with minimal

interstitial connective tissue proliferation in the birds fed with 1.0 ppm of ochratoxin.

Ravikumar (2007) observed dilatation of tubules, degeneration and desquamation

of tubular epithelium with presence of proteinaceous material in the tubular lumen and

interstitial connective tissue proliferation in birds fed with 1.0 ppm of ochratoxin.


The histological changes due to CTN toxicosis in different organs were reported by many workers,

but the target organ appears to be kidneys in almost all the species. Roberts and Mora (1978) investigated

the histopathological changes in broiler chicks fed 33, 65, 130 and 260 ppm CTN for six weeks period and

observed that all levels of dietary CTN caused cellular alternation in the kidneys. Anaplastic areas and

increased mitosis of tubular epithelial cells were reported in the kidneys. They also observed mitotic figures

in the glomeruli and epithelial lining cells of proximal tubules.

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Mehdi et al. (1981) observed degeneration and necrosis of tubular epithelial cells of both proximal

and distal convoluted tubules in broiler chicks administered with 95 mg CTN/kg body weight by crop

gavages or mixed with diet. Degenerative and necrotic changes were reported in the distal convoluted

tubules, loop of Henle and in a few proximal convoluted tubules. Acidophilic granules in the cytoplasm and

pyknotic nuclei in some of the cells were also noticed. Maryamma et al. (1990) could notice disruption of

the cytoplasmic membrane and desquamation of cells into the lumen in several foci after 4, 8 and 12 h of

feeding CTN as crop intubation at the rate of 125 mg/kg body weight. Uma and Reddy (1995) also

observed similar changes in broiler chicken fed with 125 and 250 ppm CTN for six weeks.

Ahamad and Vairamuthu (2001) observed that kidneys of birds fed with 150 and 300 ppm CTN

from 3 to 30 days of age revealed congestion, hydropic degeneration, narrowing of lumen of proximal

convoluted tubules, necrosis of tubular epithelium, individual to diffuse cellular necrosis, interstitial

fibrosis, multifocal or diffuse interstitial infiltration of mononuclear cells and a few heterophils and cellular

debris in collecting ducts. Apart from these lesions, Swaminathan (2002) observed dissociation of tubular

epithelial cells in broiler chicks fed with 150 ppm CTN up to 28 days of age.

Anandkumar (2006) observed thickening of glomerular basement membrane, degeneration and

necrosis of tubular epithelium and occasional stasis of baso-eosinophilic fluid in the lumen of tubules in

kidneys of birds fed with 5.0 ppm of citrinin.

2.13.3 Heart


Loss of cross striation, enucleation, sarcolysis, hyalinization of the cardiac fibres and mononuclear

cell infiltration between the fibres were observed when broiler chicks were fed with 125 and 250 ppm CTN

from day-old to six weeks of age (Uma and Reddy, 1995) and in broilers fed with 150 and 300 ppm CTN

from 3 to 30 days (Ahamad and Vairamuthu, 2001).

99

2.13.4 Skeletal muscle


Roberts and Mora (1978) observed skeletal myopathy resembling the myopathy seen in

hypokalaemia and vitamin E deficiency in broiler chicks fed with 62 per cent level of P. citrinum

contaminated corn.

2.13.5 Crop


Anandkumar (2006) observed hyperplasia of the crop mucosa and colonies of bacilli embedded in

the mucosa in broiler chicken fed with 5ppm CTN during 3rd and 6th week of age.

2.13.6 Proventriculus


Anitha (2007) observed increase in goblet cell activity in proventriculus of birds fed with 1.0 ppm

ochratoxin for 6 weeks.


Swaminathan (2002) observed hyperplastic changes in mucosal epithelium and mild to moderate

lymphoid infiltration in the lamina propria of proventriculus in broiler chicken fed with 150 ppm CTN

from day 1 to 28 days of age.

Anandkumar (2006) observed mononuclear cell infiltration along the base of the villi and crypt

elongation and dilation of proventricular mucosa in birds fed with 5 ppm of citrinin during 3-6 weeks of

age.

100

2.13.7 Gizzard


Anitha (2007) observed degeneration and desquamation of epithelial cells into the lumen with

increased mucus secretion in the gizzard of broiler chicken fed with 1.0 ppm ochratoxion in the diet.


Swaminathan (2002) observed interstitial fibroplasia and atrophic changes in the epithelial cells of

mucosal glands of gizzard in broiler chicken fed with 150 ppm CTN from 1 to 28 days of age.

Anandkumar (2006) observed dilatation of glands, elongation of crypts and lack of secretions,

mild interglandular fibrosis and mononuclear cell infiltration in interglandular area during 3-6 weeks of age

in broiler chicken fed with 5 ppm citrinin.

2.13.8 Intestine


Uma and Reddy (1995) observed mild catarrhal changes, focal desquamation of lining epithelium

and moderate to severe mononuclear cell infiltration in the lamina propria and mucosa and hyperplasia of

glandular epithelium of intestine in broiler chicks fed with 125 and 250 ppm CTN from day-old to six

weeks of age.

Ahamad and Vairamuthu (2001) reported mild to moderate goblet cell activity of enterocytes in

broiler chicken fed with 150 and 300 ppm CTN from 3 to 30 days of age.

Swaminathan (2002) observed loss of enterocytes in the villi with infiltration of leukocytes in the

lamina propria, which extended into submucosa and marked distension of crypts in broiler chicken fed with

150 ppm citrinin for 28 days of age.

Anandkumar (2006) observed hyperactive goblet cells and fusion of villi of intestine in broiler

chicken fed with 5ppm CTN during 3-6 weeks of age.

101

2.13.9 Pancreas


Roberts and Mora (1978) reported hydropic degeneration of acinar cells and infiltration of

lymphocytes and eosinophils in pancreas when broiler chicks were fed with 130 ppm CTN and anaplastic

cells when broiler chicks were fed with 260 ppm CTN from day 1 to 6 weeks of age and suggested that

CTN could be an allergent and carcinogen.

Ahamad and Vairamuthu (2001) reported that pancreas evinced mild vacuolar degeneration and

necrosis of acinar cells, multifocal mononuclear cell infiltration in broiler chicks fed with 150 and 300 ppm

CTN from 3 to 30 days of age. In addition to the above lesion, focal hyperplasia of the islet cells was also

observed when 150 ppm CTN was fed to broiler chicks from 1 to 28 days of age (Swaminathan, 2002).

Additionally, Anandkumar (2006) also observed reduced zymogen granules and mononuclear cell

infiltration in pancreas of broiler chicken fed with 5 ppm of citrinin for 6 weeks.

2.13.10 Lymphoid organs


Peckham et al. (1971) noticed depletion of lymphoid cells in bursa of Fabricius together with

congestion, haemorrhages and reduction in size of follicles which were often surrounded with dense fibrous

connective tissue with heterophilic and mononuclear cell infiltration in the subepithelial and interfollicular

areas in broilers fed with ochratoxin contaminated diet.

Spleen of broilers fed with 4 ppm of OA for 20 days showed marked reduction in the number of

lymphocytes in the white pulp and number of germinal centers with indistinct demarcation between the

white and red pulp in some areas. Depletion of lymphoid cells which was apparent in the cortex than in the

medulla, together with congestion, hemorrhage and heterophilic infiltration were noticed in the thymus of

birds fed with 4ppm OA for 4 wk (Dwivedi and Burns , 1984a).

102

Depletion of lymphoid elements in the bursa of Fabricius, thymus and spleen were noticed in birds

fed with ochratoxin (Ramadevi et al., 1998; Stoev et al., 1999).

Prakash (2001), Rajeev (2001) and Amita (2001) observed congestion, haemorrhages,

lymphocytolytic activity and presence of histiocytes with cellular debris in bursa of Fabricius and thymus

of broilers fed with 2 ppm of ochratoxin in the diet.

Elizabeth et al. (2002) reported that bursa of Fabricius of birds exposed to 2 ppm ochratoxin in the diet had lower number

of mitotic cells, along with lymphofollicular depletion and some pyknotic nuclei in degenerated cells.

Krisztina et al. (2002) observed lesions in spleen which included lymphocyte depletion in B-dependent and T-dependent

zones along with increased number of reticulocyte in the red pulp of spleen in experimental ochratoxicosis.

Sakthivelan and George (2002) reported depletion of lymphoid elements in the bursa of Fabricius,

thymus and spleen in broilers fed with ochratoxin levels ranging from 1 to 2 ppm.

Mild degree of congestion, focal areas of haemorrhages, degeneration of the Hassel’s corpuscles

in thymus and severe connective tissue proliferation, moderate heterophilic infiltration and pronounced

lymphocytic activity in bursa of Fabricius of broiler chicken were noticed when they were fed with 1.0 ppm

of ochratoxin A in the diet (Anilkumar, 2002 and Bhanuprakash, 2002).

Santhosh Kumar (2003) recorded congestion, haemorrhages and sparse cellularity of follicles,

lymphocytolytic activity and presence of histiocytes in lymphoid organs of birds fed with 2.0 ppm

ochratoxin in the diet for 42 days of age.

Aravind Kumar et al. (2004) observed depletion of lymphocytes and areas of congestion in thymus along with

haemorrhage in white pulp of spleen and extensive vacuolation of bursal follicles in broilers fed with ochratoxin A contaminated diets.

103

Shivappa (2005) observed lymphocytolytic activity, cellular sparcity and moderate increase in number of histiocytes in

bursa of Fabricius. In thymus, congestion, multifocal haemorrhages involving both cortex and medulla with increase in number of

histiocytes and lymphoid depletion in spleen were observed in broilers fed with 0.5 mg of ochratoxin for five weeks.

Anitha (2007) and Ravikumar (2007) observed multifocal areas of lymphocytolysis and

development of secondary lymphoid follicles in spleen, lymphocytolysis, depletion and atrophy of follicles

surrounded by fibrous tissue and micro cysts in plical epithelium of bursa of Fabricius and depletion of the

lymphoid cells, congestion and mild heterophilic infiltration in thymus of broiler chicken fed with 1.0 ppm

ochratoxion.


Lymphoid depletion and necrosis in the cortex and medulla in the bursa of Fabricius of broiler

chicks given a single dose of 47.5, 71, 100, 250 and 500 mg CTN/kg body weight were observed by Mehdi

et al. (1981) and in seven-days-old turkey poults and duckling fed 50 and 57 mg CTN/kg body weight

(Mehdi et al., 1983).

In spleen, lymphoid necrosis and depletion were characterized by shrunken white pulp and poorly

defined lymphoid sheaths in broiler chicks fed with single dose of 90, 100 and 110 CTN mg/kg body

weight. In thymus, lymphoid necrosis was pronounced in the medulla than cortex and lymphoid atrophy

was involved in thymic cortex of chicks fed with single oral dose of 47.5, 71, 100 and 500 mg CTN/kg

body weight (Mehdi et al., 1981).

In broiler chicks fed 150 and 300 ppm CTN from 3 to 30 days, bursa of Fabricius showed mild to

moderate diffuse lymphoid depletion, degeneration of lymphocytes and bursal epithelial hyperplasia.

Spleen revealed moderate lymphoid depletion with reticular cell hyperplasia. Thymus of chicks fed with

300 ppm CTN, exhibited mild depletion of cortex, but 150 ppm group did not reveal any significant change

(Ahamad and Vairamuthu, 2001).

104

Swaminathan (2002) reported lymphoid depletion in the follicles of bursa of Fabricius and

interfollicular fibrosis with infiltrating leukocytes in broiler chicks fed 150 ppm CTN for 28 days of age.

Spleen of CTN fed birds revealed mild multifocal lymphoid depletion in the germinal centres.

Anandkumar (2006) observed congestion and mild generalised lymphoid depletion,

lymphocytolysis both in the cortex and medulla in the third week and atrophic changes with severe

lymphoid depletion and corrugation of surface epithelium of bursa of Fabricius in the sixth week and

heterophilic infiltration lymphoid depletion, lymphocytolysis, mild reticular cell hyperplasia, apoptotic

bodies during third week and severe lymphoid depletion and reticular cell hyperplasia in the spleen during

sixth week in broiler chicken fed with 5 ppm citrinin.

2.14 Ultrastructural pathology


Farshid et al. (1996) reported time dependent ultrastructural lesions in the bursa of Fabricius,

spleen and thymus when pure OA was inoculated to quail embryos at the rate of 0.02 µg into the air sac of

each egg and subsequently sacrificed on day 13 and 16 after inoculation. They reported lack of

differentiation in the lymphoid cells, change in the shape of the nucleus. Granular and fibrillar structures

were present in the nucleoli. Mitochondrial change was apparent. These changes were more severe on day

16.

Santhosh Kumar (2003) reported depletion, apoptosis and necrosis of lymphocytes of bursa of

Fabricius under TEM in ochratoxin fed birds. In addition, budding with formation of apoptotic bodies and

mitochondrial damage were also observed. Further, he noticed decreased lymphocytes in the thymus with a

thin rim of cytoplasm and very few or no cytoplasmic organelles.


Brown et al. (1986) reported ultrastructurally, the normal glomarular architecture in kidneys of

layer chicks fed 300 µg CTN/g of feed from 0 to 21 days of age. They described proximal tubular epithelial

cells with ring shaped mitochondria that surrounded a central area of cytoplasm. Mitochondrial matrix

105

granules were normal in number and appearance. In some cells, peroxisomes were increased in number.

Large membrane bound 0.8 to 1.2 µm structures were present in some cells. A few proximal tubular cells

had large areas of cytoplasm filled with smooth endoplasmic reticulum. Some nuclei possessed clear 2 µm

vacuoles, partly filled with flocculent material much less dense than the cytoplasm. The authors opined

that such vaculoles might represent cytoplasmic intranuclear invaginations or invaginations of inner nuclear

membrane. Distal tubular epithelial cells possessed marginally increased numbers of cytoplasmic folds on

basal and lateral surfaces and increased numbers of large mitochondria within these folds and the collecting

duct epithelium appeared normal. Proximal tubular epithelial cells had numerous ring shaped mitochondria

enclosing areas of cytoplasm. Degenerated mitochondria were also present within the phagosomes in some

cells. Distal tubular cells infrequently possessed increased numbers of mitochondria.

Anandkumar (2006) reported vacuolations of tubular epithelial cells and swollen mitochondria

with disruption of cristae in the tubular epithelial cells. In addition, he also described the tubular cells with

large areas of cytoplasm filled with smooth endoplasmic reticulum and vacuoles in the cytoplasm and

swollen mitochondria in the liver of broiler chicken fed with 5 ppm CTN.

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New II. REVIEW OF LITERATUREshodhganga.inflibnet.ac.in/bitstream/10603/6964/6/06... · 2015. 12. 4. · ranged from 10 to 300ppb. High incidence was reported in corn byproducts (57%)