Pathological effects of feeding aflatoxin-contaminated feed on immune status and reproductive performance of juvenile white leghorn males and its mitigation with ∝-tocopherol and Moringa oleifera.

This study was planned to detect the adverse pathological consequences of aflatoxin B1 in White Leghorn (WLH) layer breeder males. Eight-week-old male layer cockerels were separated into six experimental categories: A group was kept as negative control, offered with normal feed only; group B was fed with 400 ppb amount of aflatoxin, while groups F and D fed with normal feed and supplemented with vitamin E 100 ppm and 1% Moringa oleifera, respectively, whereas groups E and C were fed with 400 ppb aflatoxin containing feed and ameliorated with vitamin E 100 ppm and 1% Moringa oleifera, respectively. This study was continued for 2 months and immunologic disorders and reproductive parameters were observed during the trial. To find out immunological status lymphoproliferative response to phytohemagglutinin-P (PHA-P), antibody titers against sheep red blood cells (SRBCs) and carbon clear assay were performed by collecting samples from five birds from each group. The whole data was measured by ANOVA test, and group means were compared by DMR test by using M-Stat C software. Regarding the reproductive status, spermatogenesis, blood testosterone level, testes weight, testes histology, sperm motility, and morphology were negatively affected by aflatoxins, but these deviations positively ameliorated by vitamin E and Moringa. Vitamin E and Moringa found advantageous in boosting the immune status of affected bird. All the immunological parameters including antibody titers against sheed red blood cells, lymphoproliferative response to avian tuberculin and phagocytic potential of macrophages were suppressed by AFB1 however in control, Moringa and vitamin E groups these immunological responses were significantly higher.

Unraveling the combined deleterious effects of ochratoxin A and atrazine upon broiler's health: Toxicopathological, serum biochemical and immunological perspectives.

Atrazine, a herbicide, is used for eradication of broad-leaved herbs in corn crop; and ochratoxins, particularly ochratoxin A (OTA), are major pollutants of poultry diet. Existence of both of these hazardous chemicals as residues is obvious as elucidated by various epidemiological findings. The present study was designed to investigate toxicopathological, serum biochemical and immunological alterations incurred by atrazine alone and/or, in combination with OTA in broilers. For this purpose, one-day old broiler chicks (n = 180) were purchased from a local hatching unit and were fed two levels of atrazine (50 and 150 mg/kg) and one level of OTA (100 µg/kg) in different combinations. Results of this experiment showed a significant reduction in feed intake, body weight gain, relative organ weights, serum total protein, albumin and globulin while there was a significant increase in urea and creatinine levels, decreased antibody response to sheep red blood cells, reduced lymphoproliferative response and phagocytic capacity in groups given OTA and atrazine individually in feed and these effects became more pronounced when atrazine was given in combination with OTA suggesting synergistic effects of both toxicants for each other.

Oxidative stress and toxicity produced by arsenic and chromium in broiler chicks and application of vitamin E and bentonite as ameliorating agents.

The present study investigated the adverse effects of arsenic and chromium in broilers and ascertained the role of vitamin E and bentonite in alleviating their harmful effects. For this purpose, we experimented on 180 one-day-old broiler chickens. The feed was administered to broiler chicks of groups 2, 6, 7, 8, and 9 chromium @ (270 mg.kg-1 BW). Groups 3, 6, 7, 8, and 9 were administered arsenic @ (50 mg.kg-1 BW). Groups 4, 7, and 9 received vitamin E (150 mg.kg-1 BW), and groups 5, 8, and 9 received bentonite (5%), respectively. Group 1 was kept in control. All the broiler chicks treated with chromium and arsenic showed a significant (p < 0.05) decline in erythrocytic parameters on experimental days 21 and 42. Total proteins decreased significantly, while ALT, AST, urea, and creatinine increased significantly (p < 0.05). TAC and CAT decreased significantly (p < 0.05), while TOC and MDA concentrations increased significantly (p < 0.05) in chromium and arsenic-treated groups on experimental days 21 and 42. Pearson correlation analysis revealed a strong positive correlation between TAC and CAT (Pearson correlation value = 0.961; p < 0.001), similarly TOC and MDA positive correlation (Pearson correlation value = 0.920; p < 0.001). However, TAC and CAT showed a negative correlation between TOC and MDA. The intensity of gross and microscopic lesions was more in chromium (270 mg.kg-1) and arsenic (50 mg.kg-1) singly or in combination-treated groups. Thus, broiler chicks treated with chromium plus arsenic exhibited higher gross and microscopic lesion intensity than other treated groups. Fatty degeneration, severe cytoplasmic vacuolar degeneration, and expansion of sinusoidal spaces were the main lesions observed in the liver. Kidneys showed renal epithelial cells necrosis, glomerular shrinkage, and severe cytoplasmic vacuolar degeneration. Co-administration of bentonite along with chromium and arsenic resulted in partial amelioration (group 8) compared to groups 7 and 9, administered arsenic + chromium + vitamin E and arsenic + chromium + vitamin E + bentonite, respectively. It was concluded that arsenic and chromium cause damage not only to haemato-biochemical parameters but also lead to oxidation stress in broilers. Vitamin E and bentonite administration can ameliorate toxicity and oxidative stress produced by arsenic and chromium.

Pathological effects of graded doses of aflatoxin B1 on the development of the testes in juvenile white leghorn males.

Current experiment was planned to investigate the deleterious effects of the graded doses of aflatoxin B1 (AFB1) on white leghorn male birds. For this purpose, one-hundred birds of 8 weeks of age were divided into 4 equal groups and reared on feed contaminated with different doses of AFB1 for 10 weeks. Group A was kept as a control group and was fed with normal toxin-free diet; groups B, C, and D were offered feed containing 100 ppb, 200 ppb, and 400 ppb of AFB1, respectively. The birds were euthanized at the 4th and 10th week of the experiment. Clinical signs, behavioral changes, absolute and relative organ weight of the testes, and sperm motility were measured. Cellular immune response was observed through carbon clearance assay (CCA), P-HAP, and antibody response against sheep red blood cells (SRBC). Results showed a dose-dependent decline in the immune response of birds with the increase in the level of AFB1 in the feed. A significant decrease in the serum levels of testosterone, prolactin, and LH were observed at the end of the study. Grossly, testicular size and volume were reduced in ABF1 fed birds, while histological examination showed moderate to severe necrosis of testicular parenchyma, with partial to complete arrest of spermatogenesis. Very few spermatozoa were found in group C, while they were almost absent in group D which was offered a diet containing 400 ppb AFB1. The motility of sperms was reduced in all treated groups except control. The abovementioned results showed that AFB1 had severe toxic effects on the reproductive and immunological parameters of WLH male birds in a dose-dependent manner.

Pattern of rifampicin resistance and gene xpert based molecular typing of tuberculosis patients in tertiary care hospitals.

Mycobacterium tuberculosis associated morbidity, mortality and drug resistance is a global health issue. The Gene Xpert is used for early diagnosis of TB and simultaneous detection of Rifampicin (RIF) resistance. We aimed to determine situation analysis of clinical TB in tertiary care hospitals of Faisalabad and to find out frequency of TB and drug resistance pattern by Gene Xpert. A total of 220 samples from suspected patients of TB were included in this study and 214 samples were detected as positive by Gene Xpert. Samples were classified on the basis of gender, age group (<30, 30-50 and >50 years), type of sample (sputum and pleural) and number of M. tuberculosis by ct value (cycle threshold). The results of present study showed high positive frequency of TB in male patients and in 30-50 years of age groups by Gene Xpert. High number of M. tuberculosis was found in low and medium category in TB patients. Out of 214 positive TB patients, rifampicin resistance was detected in 16 patients. In conclusion, our study identified that Gene Xpert is an effective approach for diagnosing TB by detection of M. tuberculosis and rifampicin resistance in <2 hours for rapid diagnosis and management of TB.

Sodium arsenite toxicity on hematology indices and reproductive parameters in Teddy goat bucks and their amelioration with vitamin C.

Arsenic is a major environmental toxicant of concern, affecting both female and male reproductive systems. The present study was conducted to investigate the toxic effects of arsenic on semen quality and reproductive hormones of Teddy bucks, an important domestic species in regions of Pakistan. In addition to arsenic, vitamin C was fed to a subset of animals to determine if there were ameliorative effects on reproductive parameters. Sixteen adult Teddy bucks were randomly divided into four experimental groups: A (control), B (arsenic 5 mg/kg BW/day orally), and C (arsenic 5 mg/kg BW/day plus vitamin C of an oral dose of 200 mg/kg BW/day). The animals in experimental group D were given only vitamin C (oral dose of 200 mg/kg BW/day). Animals were fed treated food once a day for 12 weeks. Semen quality parameters (volume, motility, count, sperm morphology, live dead ratio, sperm membrane integrity, and sperm DNA integrity) of bucks from each experimental group were evaluated on a weekly basis. Hematology and the level of arsenic in the blood were assessed every 2 weeks. Serum was collected fortnightly to measure reproductive and stress hormones (testosterone, luteinizing hormone, follicle-stimulating hormones, and cortisol). At the end of the study, all the animals were slaughtered, and the testes of all the animals were collected and evaluated for histopathology. Semen parameters in arsenic-treated bucks were significantly reduced (p < 0.05) compared with controls. Moreover, the levels of male hormones (testosterone, luteinizing hormone, and follicle-stimulating hormone) were significantly decreased in arsenic-treated animals, while cortisol was significantly increased with arsenic exposure. The histopathological lesions in the testes were present in the form of the loss of germinal epithelium and atrophy of Leydig cells. Supplementation of vitamin C however ameliorated the adverse effects of arsenic on semen quality and hormones. The histopathological lesions were also ameliorated due to vitamin C treatment. This study demonstrates that arsenic can adversely affect reproductive endpoints in Teddy goat bucks and supports the hypothesis that vitamin C is an effective treatment in arsenic-induced toxicosis. This study has high significance for Pakistan, as water contaminated with arsenic is a top health concern and is a recurring issue for both domestic animals and humans.

Evaluation of DNA vaccine encoding BCSP31 surface protein of Brucella abortus for protective immunity.

Brucellosis is a highly contagious and zoonotic disease and has a considerable impact on animal health and economy of a country, principally in Pakistan, where rural income largely depends upon livestock farming and dairy products. The disease burden is more in underdeveloped/developing countries due to the low economy and limited access to the diagnostic facilities. In Pakistan, the prevalence of Brucella abortus is very high, so it is the need of the hour to control this disease through more advanced methods. This study was designed with the aim to construct the DNA based vaccine of gene encoding antigenic surface protein (BCSP31). For this purpose, the BCSP31 gene was amplified, purified and ligated in pTZ57 R/T (cloning vector). Dubbed BCSP31-pTZ57 R/T vector was transformed into competent cells (DH5α). After plasmid extraction, the plasmid and pET-28a vector was restricted with EcoRI and BamHI. Again, ligation was done and dubbed pET-28a-BCSP31 transformed into E. coli (BL21). After expression, the protein was purified and used for evaluation of immunogenic response. The protective and immunogenic efficacy of the vaccine was evaluated in rabbits (n = 20). The rabbits were divided into four equal groups. Groups A-C were given purified protein diluted in normal saline @ 750, 1500 and 3000 µg/0.2 mL, respectively through intraconjunctival route. Group D was given 0.2 mL normal saline through intraconjunctival route. Specific immunoglobulin G (IgG) responses were measured through indirect ELISA on a weekly basis. The titer of IgG against the antigen was significantly (p < 0.05) higher in vaccinated groups A-C as compared to group D (control group) in a dose dependent manner. Moreover, log units of protection produced by DNA based vaccine in the rabbits (3.02) also indicated the protective efficacy of the DNA vaccine against B. abortus challenge. The response of this vaccine in rabbit suggested its potential effectiveness against Brucella abortus in large animals.

Pathological effects of concurrent administration of aflatoxin B1 and fowl adenovirus-4 in broiler chicks.

The current study was designed to investigate pathological effects of fowl adenovirus in broilers exposed to aflatoxin B1. Fowl Adenovirus-4 (FAdV-4) infection is remerging in all types of poultry birds in Pakistan. Poultry feed contamination with mycotoxin (aflatoxin) is another important global issue. A total of 125-day old broiler birds were divided into six equal groups. Group A served as control. B and C groups were administered with aflatoxin B1 (AFB1) 100 and 200 µg/kg feed. Group D was infected with FAdV-4, while groups E and F administered with both AFB1 (100 & 200 µg/kg) along with FAdV-4. These birds were monitored for clinical signs and mortality. Feed intake, body weight (BW), relative organ weights and gross & histopathological lesions were recorded. The highest mortality was observed in group F (FAdV-4 + AFB1 200 µg/kg feed) and the lowest mortality was observed in group B (AFB1 100 µg/kg feed). Body weights of all the groups were significantly (p < 0.05) lower as compared with control group. Relative weight of liver and kidneys in groups E and F were significantly higher as compared with control. Grossly, liver was swollen, anemic with round margins in groups D, E and F. Kidneys were also swollen with whitish areas indicating dead tissue. Microscopically intranuclear inclusion bodies were observed in group D-F. The hepatic parenchyma was also indicating necrotic changes along with vacuolar degeneration. In renal parenchyma, acute tubular necrosis was observed in groups C, E and F. It was concluded that AFB1 intoxication lead to dose dependent changes in liver and kidneys. Severity of the changes was increased in interactive groups of AFB1 with FAdV-4. Therefore, feed should be regularly monitored for AFB1 levels and day old chicks for vertically transmitted FAdV-4 to prevent losses.

Immunosuppressive effects of arsenic in broiler chicks exposed to Newcastle disease virus.

To assess the effects of prolonged exposure to arsenic (As, as arsenate) on host immune competence overall and resistance to Newcastle disease (ND) viral infection in particular, a study was carried out in broiler chicks. At 7 days of age, chicks were assigned to groups that would undergo varying vaccination, challenge, and/or As exposures; Group 1 was a control; Group 2 was to receive Newcastle disease virus (NDV) only; two groups (Groups 3, 4) were to be given As daily (50 mg/kg, by gavage) from Days 7-35 of the experiment. All groups underwent normal vaccination on Days 5, 23, and 32 against live NDV (B1 type, LaSota strain); two groups (Groups 2, 4) were challenged with field-isolated NDV at Day 24. At Days 14, 21, 28, and 35 of age, subsets of chicks in each group were evaluated. The results showed feed intake and weight gain were lower in As-treated and NDV-challenged chicks. In As-treated chicks, absolute and relative spleen weights were significantly greater, whereas those of the thymus significantly lower, over the entire 35-day period. Effects on bursa weights (absolute, relative) were only significantly reduced through Day 21. Antibody titers against ND were significantly reduced (vs. control) over the whole 35 days in birds that received As alone, but only significantly depressed through the first 21 days in birds that received As + NDV; thereafter, titers were significantly greater (in parallel with effects in birds that received NDV alone). In contrast, antibody responses to T-dependent antigen (Sheep red blood cells [SRBC]) were significantly lower in As only- and As + NDV-treated chicks throughout the study period. Among birds exposed to As (alone or with NDV), in situ phagocytic activity was elevated and cutaneous sensitivity responses decreased during the period from Day 28 to Day 35. NDV alone had spurious effects on phagocytic activity but did cause significant reductions in cutaneous sensitivity responses. It was concluded that arsenic decreased immunity in broiler chicks, thereby making them prone to ND.