Muhammad Zargham Khan

Residues of aflatoxin B1 in broiler meat: effect of age and dietary aflatoxin B1 levels.

This study describes the effect of dietary levels of aflatoxin B1 (AFB1) and age of the birds upon the residue level in liver and muscles of broiler chicks. In three different experiments broiler chicks of 7, 14 and 28 days of age were kept for 7 days on contaminated rations having 1600, 3200 and 6400 μg/kg AFB1. AFB1 residues were detected earlier in younger birds and those fed high AFB1 dietary levels. The highest residue levels in liver and muscles of young chicks fed 6400 μg/kg AFB1 was 6.97±0.08 and 3.27±0.05 ng/g, respectively. Maximum residue concentration was high in birds of young age and those kept on high AFB1 ration. After withdrawal of AF contaminated rations, residues clearance was slow and AFB1 was detectable in liver and muscles of birds for longer duration in younger birds and those fed high AFB1 dietary levels. AFB1 residues in poultry tissues may buildup to high levels in areas with no regulatory limits on AFB1 levels of poultry feed and may pose a risk to consumers health.

Effects of cypermethrin on some clinico-hemato-biochemical and pathological parameters in male dwarf goats (Capra hircus).

This study was carried out on 30 dwarf bucks to determine the effects of cypermethrin (CY) on clinical, hemato-biochemical and histopathological parameters. Animals were divided randomly into five equal groups, and each group was dipped in 0%, 0.1%, 0.4%, 0.8% or 1.6% CY, on days 0 and 15. Animals were monitored for clinical signs. Blood and serum samples were collected on day 0 and then fortnightly till day 75. Severe clinical signs comprising itching, restlessness, salivation, skin scratching and head shaking appeared at high doses (0.8% and 1.6% CY). Erythrocyte counts, hemoglobin, hematocrit, total protein, globulin and fibrinogen decreased significantly while total leukocyte counts, alanine aminotransferase and aspartate aminotransferase concentration increased significantly in all the treated groups. In the liver, necrosis of hepatocytes along with cytoplasmic vacuolation and fibroblasts proliferation were observed at a high dose of CY (1.6%). Microscopically kidneys showed congestion of parenchyma and condensation of epithelial cells of tubules along with deposition of casts in tubules. Shrinkage of glomerular capillaries and increased urinary spaces were pronounced in the high-dose group. Lungs exhibited accumulation of fibrinous exudation, thickening of alveolar walls, collapse and broken alveoli in animals treated with a high dose of CY. It was concluded that CY caused dose-dependent effects on all parameters studied. High doses of CY (0.8% and 1.6% solution) affected the parameters on erythrocytes and leukocytes for whole evaluation period, while effects on plasma proteins were transient and on ALT, AST and fibrinogen were transient but lasted a few weeks longer.

Toxico-pathological changes induced by cypermethrin in broiler chicks: their attenuation with Vitamin E and selenium.

Ninety 1-day old broiler chicks of mixed gender (as hatched) procured from a local hatchery were randomly divided into five equal groups. All the treatments were given through crop tubing. Groups 1-4 received cypermethrin (CY) (600mgkg(-1)b. wt.) daily for 30 days. In addition to CY (group 1), groups 2-4 received Vit E (150mgkg(-1)b. wt.), Se (0.25mgkg(-1)b. wt.), and Vit E (150mgkg(-1)b. wt.)+Se (0.25mgkg(-1)b. wt.), respectively. Group 5 served as control andreceived normal saline (2mlkg(-1)b. wt.) for 30 days. Randomly selected six broiler chicks from each group were slaughtered at experimental days 10, 20 and 30 for the collection of serum/plasma and morbid tissues. Absolute organ weights were recorded. Total plasma proteins, fibrinogen and creatinine were significantly (P<0.05) increased while alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and urea decreased significantly (P<0.05) in CY-treated group when compared with the control group. Kidneys were swollen grossly in treated broiler chicks. In liver, necrosis of hepatocytes, cytoplasmic vacuolation, bile duct hyperplasia and mononuclear cellular infiltration were observed. In kidneys, necrosis of tubular epithelial cells, cytoplasmic vacuolation, cellular infiltration and atrophy of glomeruli were observed. Sub-arachnoid space was much dilated in CY-treated broiler chicks. It can be concluded that CY induces biochemical and histopathological alterations in broilers chicks; however, these toxic effects can be ameliorated by Vit E or Se. Combination of Vit E and Se was more effective in ameliorating toxic effects of cypermethrin in broilers chicks.

Clinico-hematological and micronuclear changes induced by cypermethrin in broiler chicks: Their attenuation with vitamin E and selenium

This study was carried out on 90 one-day-old broiler chicks to know clinico-hematological alterations, DNA damage caused by cypermethrin (CY), and attenuation of toxic effects by vitamin E (Vit E) and selenium (Se). Birds were randomly divided into five equal groups. Groups 1-4 received CY (600mlkg(-1)b.wt) daily for 30 days by crop tubing. In addition to CY, groups 2, 3 and 4 received Vit E (150mgkg(-1)b.wt), Se (0.25mgkg(-1)b.wt), and Vit E (150mgkg(-1)b.wt)+Se (0.25mgkg(-1)b.wt), respectively. Group 5 served as control. Birds were monitored twice daily for clinical signs. They were weighed and blood samples were collected at experimental days 10, 20 and 30 for hematological studies. CY-treated birds showed more prominent signs of toxicity compared to CY+Vit E, CY+Se and CY+Vit E+Se birds. Body weight in groups 1-3 was significantly (P<0.05) smaller at days 20 and 30 when compared with the control group. Significantly (P<0.001) higher numbers of micronuclei appeared in chicks treated with CY compared to CY+Vit E- and CY+Se-treated birds. Significantly decreased total erythrocyte counts (TEC), hemoglobin (Hb) concentration and packed cell volume (PCV) in all treated groups were recorded. Treated birds suffered from macrocytic hypochromic anemia. Leukocytosis in early stage and later leucopenia was seen in treated birds. It can be concluded that CY induces toxic effects in broilers chicks; however, these toxic effects can be ameliorated by Vit E or Se. Combination of Vit E and Se was more effective to ameliorate toxic effects of cypermethrin.

Amelioration of Ochratoxin A-induced immunotoxic effects by silymarin and vitamin E in White Leghorn cockerels

Silymarin (SL) is the bioactive extract of the plant Silybum marianum and Vitamin E (VE) is an important anti-oxidant. The present study was designed to evaluate potential ameliorative effects of SL and VE against Ochratoxin A (OTA)-induced immunotoxic effects in White Leghorn cockerels. One day-old birds were divided into 12 groups (20 birds/group) and fed basal diets amended with OTA (1.0 or 2.0 mg/kg) alone or in combination with SL (10 g/kg) and/or VE (200 mg/kg) for 42 days. Immunological in situ responses, including antibody formation against sheep red blood cells (7 and 14 days after both primary and booster injections), lymphoproliferative responses to avian tuberculin (30 days of age), and mononuclear phagocytic system function (i.e. by clearance of injected carbon particles) assay (42 days of age), were assessed. Results suggested that silymarin and Vitamin E alone or in combination ameliorated the immunotoxic effects induced by 1.0 mg OTA/kg but could not significantly impact on the effect from ingestion of 2.0 mg OTA/kg. The results of the present study suggested that both SL and VE possess an ability to ameliorate OTA-induced immunotoxicity in chicks. However, it remains to be determined whether/what SL:OTA or VE:OTA ratios are required to assure such mitigation of OTA-induced immunotoxicities.

Effects of individual and combined administration of ochratoxin A and aflatoxin B1 in tissues and eggs of White Leghorn breeder hens

BACKGROUND Mycotoxins, the secondary fungal metabolites, are unavoidable contaminants of human and animal food and feeds. The objectives of this study were to evaluate the effect of concurrent feeding of ochratoxin A (OTA) and aflatoxin B(1) (AFB(1) ) to breeder hens, upon their deposition in different tissues and eggs. RESULTS Residues of OTA and AFB(1) in (ng g(-1) ) were significantly higher in liver followed by kidneys and breast muscles by 22.54 ± 1.48, 4.22 ± 0.93 and 0.56 ± 0.06 for OTA (group fed OTA at 5 mg kg(-1) diet) and 1.44 ± 0.21, 0.25 ± 0.01 and 0.03 ± 0.01 for AFB(1) (group fed AFB(1) at 5 mg kg(-1) diet), respectively. Residues of OTA and AFB(1) in eggs appeared at days 3 and 5 of toxin feeding and disappeared at days 5 and 6 of withdrawal of mycotoxins contaminated feed, respectively. The residues of OTA and AFB(1) were significantly lower in the tissues of hens fed these toxins concurrently compared with the groups fed OTA and AFB(1) independently. CONCLUSIONS Residues of OTA and AFB(1) appeared in the tissues and eggs of laying hens kept on OTA- and AFB(1) -contaminated diets. Concurrent feeding of OTA and AFB(1) to hens significantly decreased the concentration of OTA and AFB(1) residues in the tissues and eggs.

Toxico-Pathological Effects of In Ovo Inoculation of Ochratoxin A (OTA) in Chick Embryos and Subsequently in Hatched Chicks

This study was designed to investigate the toxico-pathological effects of in ovo inoculation of ochratoxin A (OTA) in chicken embryos and subsequently in the hatching chicks. Nine hundred fertile white leghorn (WL) layer breeder eggs were divided into eight groups (A–H). Group A was maintained as untreated control, whereas group B was kept as sham control (10 µL of 0.1 M NaHCO3 solution). Before incubation, groups C, D, E, F, G, and H were injected with 0.01, 0.03, 0.05, 0.10, 0.50, and 1.00 µg OTA/egg, respectively. At 53 hrs of incubation, crown to rump length, optic cups, and eye lens diameters were significantly (p ≤ .05) lower, whereas neural tube closure defects were higher in the OTA-treated embryos. Teratogenic defects (studied at day 9 of incubation) and embryonic mortalities were higher in the groups administered high doses of OTA. A significant increase was noted in the serum concentration of ALT, urea, and creatinine, along with higher weights of liver and kidney, in chicks hatched from OTA-contaminated eggs. These findings suggested that there are teratogenic and substantive toxicological risks in the developing chicken embryos and hatched chicks that could be exposed to OTA in ovo.

Immunological status of the progeny of breeder hens kept on ochratoxin A (OTA)- and aflatoxin B1 (AFB1)-contaminated feeds

This study aimed to evaluate the immunological status of progeny of hens kept on ochratoxin A (OTA)- and aflatoxin B1 (AFB1)-contaminated feed. For this purpose, White Leghorn (WL) layer breeder hens (40-weeks-of-age) were divided into six groups (A–F). Hens in Group A were fed a commercial layer ration while those in Groups B and C were kept on a diet amended with 3 and 5 mg OTA/Kg, respectively. Group D was fed a ration containing 5 mg AFB1/Kg, while hens in Groups E and F were kept on feed amended with OTA and AFB1 each. All feedings were for 1, 2, or 3 weeks. Fertile eggs were set for hatching on a weekly basis to obtain progeny of each week separately. At 14 days-of-age, subsets of progeny were euthanized and the frequency of immunoglobulin(s)-bearing cells in their spleen and bursa of Fabricius assessed; at 16 days-of-age, other chicks in each set were utilized to determine their lymphoblastogenic responses against phytohemagglutinin (PHA-P). At 30 days-of-age, the final sub-set of chicks/group was euthanized and their peritoneal macrophages harvested for measurements of phagocytic potential and nitrite production. Relative weights of the bursa of Fabricius and of the spleen were significantly lower in the progeny of hens fed mycotoxin-contaminated diets for 14 and 21 days. The frequencies of IgA-, IgG-, and IgM-bearing cells were also significantly lower in the bursa of Fabricius and spleen of progeny chicks obtained from hens fed the OTA + AFB1 mixed diet. Feeding contaminated diets to breeder hens also resulted in significantly lower responses to PHA-P. In addition, the percentages of peritoneal macrophages displaying phagocytosis of sheep red blood cells (SRBC), the number of SRBC/macrophage, and nitrite production were each significantly lower in cells from progeny chicks from OTA- and AFB1-fed hens. The findings of the present study indicated there were severe immunosuppressive effects in progeny chicks as a result of exposure of their parent hens to OTA and AFB1 either alone or in combination. These studies provide emphasis for the need for mycotoxin regulation policy with respect to the ingredients used in poultry feed, since it is clear that feeding multi-mycotoxin-contaminated diets to breeder hens will almost certainly result in the hatching of manifestly unhealthy chicks.

Toxicopathological effects of lambda-cyhalothrin in female rabbits (Oryctolagus cuniculus).

The aim of this study was to investigate effects of lambda-cyhalothrin (LCT) on clinical, hematological, biochemical and pathological alterations in rabbits (Oryctolagus cuniculus). New Zealand white female rabbits (n = 24) of 4-5 months age having 997.92 ± 32.83 g weight were divided into four equal groups. Group A (control) received normal saline intraperitoneally (ip). Animals in groups B, C and D were treated with LCT 1.0, 4.0 and 8.0 mg/kg bw ip. Each group received seven consecutive doses at an interval of 48 hours. Blood and serum samples were collected at an interval of 96 hours. Blood analysis revealed a significant (p < 0.05) decrease in red blood cell and white blood cell counts, hemoglobin concentration and lymphocytes, while mean corpuscular hemoglobin concentration, mean corpuscular volume, neutrophils, monocytes and eosinophils were increased. Serum biochemical analysis revealed significant (p < 0.05) decrease in serum total proteins and serum albumin, while an increase was seen in serum alanine aminotransferase and aspartate aminotransferase activities compared with the control group. Serum globulin values varied non-significantly in all treatment groups as compared to control group. A dose-dependent increase in the incidence of micronucleated polychromatic erythrocyte was observed. All gross and histopathological lesions observed in LCT-treated rabbits were dose-dependent. Liver of the treated rabbits exhibited extensive perihepatitis, hyperplasia of bile duct, necrosis, hemorrhages and congestion. In lungs, there were hemorrhages, thickened alveolar walls, congestion, emphysema, collapsed alveoli and accumulation of extensive inflammatory cells. Kidneys were congested and hemorrhagic whereas renal parenchyma and stroma were normal. Microscopically, heart showed congestion of blood vessels and nuclear pyknosis, myodegeneration. It was concluded from the study that LCT produced toxicopathological alterations in rabbits in a dose-dependent manner. On the basis of the results, it can be suggested that overdosing of LCT be avoided while treating animals for ectoparasites.

Immunological responses of male White Leghorn chicks kept on ochratoxin A (OTA)-contaminated feed

This study was designed to evaluate some immunological responses of male White Leghorn (WL) chicks kept on an ochratoxin A (OTA)-contaminated diet. For this purpose, 350 1-day-old male WL chicks were divided into five groups (A–E). Group A was kept as control, while Groups B, C, D, and E were fed OTA-contaminated feed at 0.1, 0.5, 1.0, and 1.5 mg/Kg diet, respectively, for 21 days, and then basal ration for the remaining period. At 14- and 16-days of age, random chicks (n = 10) from each group were used for analyses of phagocytic function of the reticuloendothelial system or for measuring the lymphoproliferative responses to intradermally-administered T-cell mitogen, phytohemagglutinin-P (PHA-P), respectively. At 30-days of age, abdominal macrophages were collected from 15 chicks/group and utilized for determination of their phagocytic potential and for nitrite production. Antibody (Ab) titers (i.e., total antibodies, IgM, and IgG) against sheep red blood cells (SRBC) were determined at 7 and 14 days after a primary (at 7 days of age) and a booster (given 14 days after primary [at 21-days of age]) dose (intravenous) of the antigen. Data from the present study showed that the relative weight of the bursa of Fabricius of chicks fed OTA for 14 and 21 days and the spleen of chicks fed OTA for 21 days were significantly lower than their control counterpart. Phagocytic function of reticuloendothelial system evaluated by carbon clearance, and lymphoproliferative response to PHA-P, of chicks kept on OTA-contaminated diet were significantly lowered. The percentage of abdominal macrophages displaying phagocytosis of SRBC, the number of SRBC/macrophage, and nitrite production were each significantly lower in cells from chicks in the OTA-fed groups. Total Ab (at days 7 and 14 post-booster SRBC injection) and IgG (at day 14 post-primary and day 7 post-booster SRBC injection) titers against SRBC showed significant reductions in the groups fed OTA-contaminated diet. The findings of this study are in line with the previous work suggesting the immunosuppressive effect of OTA in male WL chicks regarding functional impairment in some of the components of the immune system.