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Gentamicin is an effective and economical drug used to control infectious diseases in poultry but is highly toxic and had slow clearance from the body. This study aimed to report three cases of gentamicin toxicity in three White Leghorn (WLH) layer flocks in different poultry producing areas of Pakistan. In first case, gentamicin was injected in a 9000 WLH layer flock @ 10 mg/kg body weight (BW) for seven times during 9-15 weeks for age. In second case, gentamicin was injected in a flock of 7500 WLH layers @ 25 mg/kg BW for four times during 17-18 weeks of age. In third case, gentamicin was injected in flock of 16000 WLH layers @ 22.22 mg/kg BW three times in 20-21 weeks of age. Flock wise mortality was 8.69, 82.63 and 71.86%, respectively. Birds were dehydrated, emaciated and had prominent keel bone. Clinical signs included dehydration, decreased body weight leading to emaciation, decreased feed intake, increased water intake and watery diarrhea. Necropsy revealed prominent keal bone, shrunken muscles swollen kidneys bulging out from bony sockets. Petechial and echymotic hemorrhages were present on heart and skeletal muscles. Liver was enlarged with hemorrhagic streaks on its surface. Microscopically, hemorrhages and acute tubular necrosis was recorded in kidneys. Liver had hemorrhages, cellular infiltration and vacuolar (fatty) degeneration of hepatocytes. From the results, it could be concluded that overdosing and repeated administration of gentamicin was highly toxic to birds. [ABSTRACT FROM AUTHOR]

The present study was carried out to find toxico-pathological effects of cottonseed meal (CSM) in male Japanese quail (Coturnix japonica). Male birds (n=48) were equally divided into four groups (A to D). Three isonitric and isocaloric experimental feeds were formulated by replacing soybean meal with three levels of CSM i.e., 13, 27 and 41%. The respective feed was offered to the birds ad libitum for the duration of experiment (42 days). Clinical signs, behavioral alterations, feed consumption, body weight, absolute and relative organ weight, hematological and biochemical parameters along with gross and histopathological lesions were studied. In group B and C, birds were temporarily depressed but later on became active. In group D, birds remained dull and depressed and 66.7% mortality was recorded. Body weight, absolute and relative organ weight was non-significantly different in treatment groups compared with control. Feed intake at week 1 was significantly low in group D while during remaining experiment; it differed non-significantly in all the treatment groups compared with control. Testicular volume at day 21 was significantly (P<0.05) low in group D. Significantly low serum total proteins and albumin in groups B and C and hematocrit values in all the groups and hemoglobin concentration in group D were recorded at day 42 of experiment. It was concluded that CSM 13% level did not have any deleterious effect on the feed conversion and body weight but the reproductive performance of the male Japanese quail was affected. [ABSTRACT FROM AUTHOR]

Lindane is a broad-spectrum insecticide widely used on fruits, vegetables, crops, livestock and on animal premises to control the insects and pests. The extensive use of pesticides and their residues in the soil and water typically join the food chain and thus accumulate in the body tissues of human and animals causing severe health effects. The study was designed to determine the toxicity effects of sub-lethal concentrations of lindane on hemato-biochemical profile and histo-pathological changes in Rohu (Labeo rohita). A significant increase in the absolute (p<0.05) and relative (p<0.05) weights was observed along with severe histo-pathological alterations in liver, kidneys, gills, heart and brain at 30μg/L and 45μg/L concentration of lindane. A significant (p<0.05) decrease in RBCs count, PCV and Hb concentration while a significant (p<0.05) increased leukocytes were observed by 30μg/L and 45μg/L concentrations of lindane at 45 and 60 days of the experiment. Serum total protein and albumin were significantly (p<0.05) decreased while hepatic and renal enzymes were significantly (p<0.05) increased due to 30μg/L and 45μg/L concentrations of lindane at days-45 and 60 of experiment compared to control group. The observations of thin blood smear indicated significantly increased number of erythrocytes having nuclear abnormalities in the fish exposed at 30μg/L and 45μg/L concentrations of lindane. ROS and TBARS were found to be significantly increased while CAT, SOD, POD and GSH were significantly decreased with an increase in the concentration and exposure time of lindane. The results showed that lindane causes oxidative stress and severe hematological, serum biochemical and histo-pathological alterations in the fish even at sub-lethal concentrations., Competing Interests: The authors declare no conflict of interest., (Copyright: © 2024 Afzal et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Spermatozoa cryopreservation has been practiced for decades and is a very useful technique for long-term preservation of sperm fertility. The capability for semen cryopreservation varies across species, seasons, latitudes, and even for different ejaculates from the same animal. This article summarizes research results on sperm cryotolerance biomarkers in several species, focusing on three areas: spermatozoa cryotolerance biomarkers, seminal plasma proteins cryotolerance biomarkers, and other cryotolerance biomarkers. We discovered that sperm cryoresistance biomarkers are primarily related to sperm plasma membrane stability, the presence of antioxidant substances in sperm or seminal plasma, sperm cell energy metabolism, water and small molecule transport channels in the sperm plasma membrane, and antistress substances in sperm or seminal plasma. The research conducted using diverse livestock models can be employed to enhance the basic and applied reproduction of other mammals through the study of sperm cryotolerance biomarkers, as well as the substantial similarities between livestock and other organisms, including endangered species., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Suo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The largest monkeypox virus (MPXV) outbreak of the 21st century occurred in 2022, which caused epidemics in many countries. According to WHO, physical contact with infected persons, contaminated surfaces, or affected animals might be a source of this virus transmission. A febrile sickness including few symptoms found in MPX disease. Skin rash, lesions, fever, headache, fatigue, and muscle aches symptoms were observed commonly for this disease. Animal and in vitro, studies have shown that the antiviral medications cidofovir and brincidofovir are effective against MPXV. The first-generation vaccinia virus vaccine was developed in 1960, and it helped to protect against MPXV with its side effects. A second-generation vaccination with limitations was launched in 2000. However, the CDC advised vaccinations for risk groups in endemic countries, including positive patients and hospital employees. The JYNNEOS vaccine, administered in 2 doses, also provides protection from MPX. This article presents concisely the most recent findings regarding epidemiology, genomic transmission, signs and symptoms, pathogenesis, diagnosis, and therapeutic interventions for MPXV, which may be helpful to researchers and practitioners. WHO declared that MPX was no longer a global health emergency due to its declining case rate, and a number of countries have reported new incidences. Further research-based investigations must be carried out based on the 2022 outbreak., (© 2023 The Author(s).)

Despite being an essential trace element for numerous metabolic processes and micronutrients, copper (Cu) has induced adverse effects on the environment and public health due to its continuous and widespread use for the last several decades. The current study assessed the hematological and histopathological alterations in the freshwater fish ( Labeo rohita ) exposed to graded concentrations of copper sulfate. For this purpose, L. rohita fish ( n = 72), weighing ~200-215 g, were randomly divided into four experimental groups and then exposed to acute doses of CuSO 4 , i.e., control, 0.28, 0.42, and 0.56 μgL -1 . For comparative analysis of hematological and biochemical changes, blood/serum samples were obtained on 12, 24, and 36 days. Overall, the body weight of fish decreased with the time and dose of CuSO 4 ; as the dose increases, body weight decreases. Dose and time-dependent results were observed in other parameters also. Results showed a significant increase in leukocytes, whereas red blood cells count, Hb, and Hct were significantly reduced in treated groups compared to the control. The mean corpuscular hemoglobin (MHC) and mean corpuscular hemoglobin concentration (MCHC) showed a non-significant decrease in treated groups compared to the control group. Serum biochemical parameters, including total proteins, albumin, and globulin, decreased significantly ( p < 0.05). At the same time, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), glucose, and cholesterol were significantly ( p < 0.05) increased in the treated groups compared to the control group. Significantly ( p < 0.05) increased levels of lipid peroxidation while decreased values of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (RGSH) in the blood of fish were recorded. Histopathological examination of fish gills, liver, and kidneys showed inflammation and degenerative changes due to CuSO 4 exposure. In the brain tissue, degenerative changes like neuron necrosis, intracellular edema, cytoplasmic vacuolization, and congestion were observed. In conclusion, the study indicates that exposure to copper sulfate, even in smaller concentrations, can cause adverse hematological and histopathological changes in L. rohita fish., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Naz, Hussain, Guangbin, Chatha, Rehman, Jahan, Liaquat and Khan.)

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., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Mashkoor, Al-Saeed, Guangbin, Alsayeqh, Gul, Hussain, Ahmad, Mustafa, Farooq and Khan.)

Rheumatoid arthritis (RA) is an inflammatory condition and associated with the symmetrical synovitis of the joints and cause joint pain. The use of anti-rheumatic drugs is associated with many adverse effects. Quercetin, an important polyphenolic flavonoid, possess anti-inflammatory and anti-rheumatic effects. Quercetin use is limited due to poor absorption and bioavailability. Nanomedicines are used for the targeted drug delivery, hence it reduces the adverse effects of the drug. Based upon these factors, quercetin-loaded chitosan nanoparticles (Q-NPs) were prepared by solvent evaporation method, characterized and their better anti-rheumatic effect with mechanistic insights was validated in Freund's complete adjuvant (FCA)-induced arthritic rats along with safety studies. The animals were divided into five groups, each containing 5 animals. Group I was normal control, group II was arthritic control, while groups III, IV and V were administered with quercetin (15 mg/Kg) and Q-NPs (10 and 20 mg/Kg), respectively. The reduction in ankle diameter, serum oxidative stress markers as well as pro- and inflammatory cytokines, e.g., tumor necrosis factor (TNFα), interleukin (IL-6) were determined. The prepared Q-NPs showed hydrodynamic size of 83.9 nm, polydispersity index of 0.687, entrapment efficiency 90.5% as well as no interaction between quercetin and chitosan in Fourier transform infrared spectroscopy (FTIR). A significant reduction (p < 0.001) in ankle diameter was observed after administration of high-dose Q-NPs (4.32 ± 0.14 cm to 5.13 ± 0.62 cm). There was also reduction (p < 0.001) in levels of TNFα and IL-6 following high-dose Q-NPs (72.56 ± 2.30 and 308.19 ± 11.5 pg). The effect on biochemical tests, hematological parameters and oxidative stress parameters was also found to be significant. Histopathological changes of kidney, liver and ankle also confirmed the anti-rheumatic effect of high-dose Q-NPs. The study concludes that administration of Q-NPs (20 mg/Kg) may be used for the treatment of FCA-induced RA in rats., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

This study was designed to compare immunopathological effects of in ovo vaccination with post-hatch vaccination against IBD in White Leghorn chicks. A total of 189 embryonated eggs were divided into six groups. At day 18 of incubation, groups A-C were administered in ovo with 228E, Winterfield 2512:10/3 and 2512/90:10/2.7, respectively, group D (post-hatch vaccination) and group E as shamed control (for quality evaluation of in ovo vaccination technique), and group F as control. The results showed that antibody titers against IBD detected by ELISA on days 2, 17, and 28 were significantly higher in all in ovo groups as compared to control groups E and F. On day 17, all vaccinated groups ( in ovo and post-hatch vaccinated) showed no significant differences in antibody titers among themselves; however, at day 28, only the post-hatch group showed significantly higher antibody titers followed by in ovo vaccinated groups. The cell-mediated immunity determined by PHA-P assay was significantly higher in all vaccinated groups than the non-vaccinated groups. No clinical signs of IBD infection were observed in any of the vaccinated groups. There was only increase in bursa size of groups vaccinated with intermediate plus strains (groups A, C, and D) at day 28. The histopathology showed that all the treatment groups had mild lesions induced by IBD virus in bursa. This study concluded that in ovo vaccination with live IBD vaccines provides protective immunity to the chickens even in the presence of IBD-specific MDA; therefore, the onset of immunity was much earlier than the post-hatch vaccination and in ovo groups also maintained protective immunity against IBD for longer time., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zaheer, Chen, Khan, Elokil, Saleemi, Zaheer and Khan.)

Brucellosis is a well-known and harmful zoonotic disease that poses a severe threat to public health and wild and dairy animals. Due to a lack of monitoring and awareness, disease incidence has increased. Therefore, this study was conducted for the first time to ascertain the status of seroprevalence of brucellosis, hematological, oxidative stress, and antioxidant enzymes in different breeds of cattle reared under tropical-desert conditions in Pakistan. This study comprised 570 cattle of different breeds. We recorded some epidemiological traits, including age and gender. The blood samples were obtained from all the cattle, screened with RBPT, and then confirmed by ELISA and PCR. The results recorded an overall 11.75%, 10.7%, and 9.64% prevalence of brucellosis based on RBPT, ELISA, and PCR. We obtained nonsignificant results in different age and sex groups of cattle. The results showed significantly ( P ≤ 0.05) lower values of erythrocyte counts, hemoglobin quantity, hematocrit, lymphocytes, and monocytes in infected cases. The results showed that the total leukocyte and neutrophil cells significantly ( P ≤ 0.05) increased. The lipid peroxidation parameters (MDA- and NO-scavenging activity of erythrocyte) increased significantly ( P ≤ 0.05) in infected cattle, whereas significantly reduced antioxidant enzymes like SOD, RGSH, and CAT were. Similarly, significantly lower serum albumin levels and total serum proteins were recorded in infected cattle., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2022 Riaz Hussain et al.)

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., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Pyriproxyfen (PPF) mimics a natural hormone in insects and disrupts their growth. It is a well-known synthetic insecticide and aromatic juvenile hormone analog frequently used in agriculture and vegetable crops to control various insect species. At present, scanty information is available about the possible potential threats of PPF in aquatic organisms. Therefore, in this study, different toxico-pathologic endpoints of PPF like DNA damage, biomarkers of oxidative stress, and status of antioxidant enzymes were determined in Labeo rohita (freshwater fish). In our study, 60 active, free from any external obvious ailments, same size, age, and body mass were randomly allocated to four glass aquaria (T0-T3) separately containing 100 L water. The fish present in groups T1, T2, and T3 were administered PPF dissolved in water 300, 600, and 900  μ g/L for 30 days. Different tissues including the blood and visceral organs were obtained from each fish on days 10, 20, and 30 of the experiment. Results on various morphological and nuclear changes in red blood cells of PPF-exposed Labeo rohita fish including pear-shaped erythrocytes, spherocytes, red blood cells with a blebbed nucleus, micronucleus, and nuclear remnants were significantly increased. Our results on genotoxicity (comet assay) recorded significantly ( P ≤ 0.05) increased DNA damage in various tissues of insecticide-exposed fish. The results on oxidative stress profile (reactive oxygen species and thiobarbituric acid reactive substances) and antioxidant enzymes (reduced glutathione superoxide dismutase, peroxidase, and catalase) in multiple tissues of Labeo rohita fish concluded significantly ( P ≤ 0.05) higher quantity of biomarkers of oxidative stress and lower concentrations of different antioxidant enzymes in treated fish. Hence, the findings of our experimental research determine that PPF could induce adverse toxic impacts on multiple tissues of Labeo rohita fish., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2022 Xuwen Li et al.)

Mycobacterium bovis ( M. bovis ) being the main cause of animal tuberculosis is a complex infectious agent and can be a cause of zoonotic tuberculosis zoonosis in public health. To date, the uncommon infection in public health due to M. bovis still is a great challenge to both veterinary and medical professions and requires a careful diagnosis and confirmation of the bacterium. Therefore, this study for the first time reports the clinical, gross, histopathological, and molecular based confirmation of M. bovis infection in wildlife animals (nilgai). Prior to death, the morbid animal showed severe pneumonic ailments like moist cough, thick nasal exudates, and dyspnoea. At necropsy, enlargement of mandibular cervical and mesenteric lymph nodes was observed. Different macroscopic lesions such as congestion and hyperaemia, creamy white and catarrhal exudates in trachea, consolidation, grey and red hepatisation of lungs, and micro- and macrogranulomatous tubercles containing caseous materials in lungs were observed. The heart of morbid animal showed congestions, myocarditis, and a copious amount of straw-colored fluid in the pericardial sac. At the microscopic level, lungs indicated granulomatous inflammatory response, presence of multinucleated giant cells, fibrosis, and punctuation of alveoli with chronic inflammatory cells. Histopathological examination of various sections of the heart of the infected animal showed chronic inflammatory response consisting of chronic inflammatory cells like monocyte, lymphocytes, and fibroblasts along with noncalcified eosinophilic materials. At the molecular level, M. bovis infection was confirmed in various tissues like the heart, lungs, cervical, and mesenteric lymph nodes in morbid animals. In conclusion, based on our results, it can be suggested that more molecular based epidemiological studies are crucial to know the exact cause of pulmonary and cervical tuberculosis in wild animals., Competing Interests: The authors declare no conflict of interest., (Copyright © 2022 Riaz Hussain et al.)

Acetochlor is persistently used in the agroproduction sector to control broadleaf weeds. Due to frequent and continuous applications, this herbicide can reach nearby water bodies and may induce deleterious changes in aquatic life. Therefore, investigation of harmful impacts of different environmental pollutants, including herbicides, is vital to knowing the mechanisms of toxicity and devising control strategies. The current experiment included bighead carp ( n = 80) to estimate adverse impacts. Fish were randomly placed in 4 different experimental groups (T0-T3) and were treated for 36 days with acetochlor at 0, 300, 400, and 500  μ g/L. Fresh blood without any anticoagulant was obtained and processed for nuclear and morphological changes in erythrocytes. At the same time, various visceral organs, including the gills, liver, brain, and kidneys, were removed and processed on days 12, 24, and 36 to determine oxidative stress and various antioxidant biomarkers. Comet assays revealed significantly increased DNA damage in isolated cells of the liver, kidneys, brain, and gills of treated fish. We recorded increased morphological and nuclear changes ( P ≤ 0.05) in the erythrocyte of treated fish. The results on oxidative stress showed a higher quantity of oxidative biomarkers and a significantly ( P ≤ 0.05) low concentration of cellular proteins in the gills, liver, brain, and kidneys of treated fish compared to unexposed fish. Our research findings concluded that acetochlor renders oxidative stress in bighead carp., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2022 Yasir Mahmood et al.)

Mycobacterium bovis causes tuberculosis in dairy and wild animals. Presence of tuberculosis in animals poses a threat not only to their herd mates but also for public. No reports are available about the clinical, pathological, and molecular investigation of naturally occurring tuberculosis (TB) due to M. bovis in one-horned rhinoceros. One-horned female rhinoceros ( Rhinoceros unicorns ) at the age of 41 years died in a public park in Pakistan. Postmortem and other investigations were carried out to know the cause of death. The present study describes necropsy, histopathology, and molecular-based confirmation of TB in a captive female rhinoceros that died of this infection. Clinically, the rhinoceros showed nonspecific clinical signs including anorexia, lethargy, dyspnoea, coughing, and sudden death. At necropsy, the trachea exhibited mild congestion and contained catarrhal exudate at the bronchial bifurcation. Macroscopic examination revealed characteristic tubercles on all parenchymatous organs. The lungs showed consolidation, grey hepatization, and contained granulomatous lesions packed with cheesy exudate. Histopathological examination showed severe pneumonic changes in the form of granulomatous inflammation consisting of lymphocytes, multinucleated giant cells, caseous materials, and mineralized foci surrounded by a fibrous capsule. PCR amplicon of 500 bp confirmed the presence of M. bovis in multiple hepatic and pulmonary tissue samples, as well as in uterine exudates. It was concluded that the presence of tuberculosis in rhinoceros may pose potential transmission risk to other animals and the application of practical tools to determine TB status in the rhinoceros is crucial., Competing Interests: The authors declare no conflict of interest., (Copyright © 2022 Abu Baker Siddique et al.)

Currently, aquatic and terrestrial ecosystems are continuously and chronically polluted by cocktails of countless chemical compounds. The susceptibility to infections is tremendously increasing in a variety of organisms due to exposure to environmental pollutants. Pendimethalin, an herbicide, is continuously used in agriculture to remove unwanted broadleaf weeds across the globe. Therefore, this study investigates the mechanisms of toxicity of pendimethalin in freshwater fish bighead carp upon exposure to low and environmentally relevant concentrations. For this purpose, 48 fish without any clinical abnormalities were kept in a glass aquarium in different experimental groups (T0, T1, T2, and T3). These groups were treated with pendimethalin at 0.00, 0.25, 0.50, and 0.75 mg/L, respectively. Four fish were randomly picked from each experimental group and killed at 72, 96, and 120 hours of the trial to study hematobiochemical parameters and visceral tissues including the brain, liver, heart, gills, and kidneys for histopathology. Herbicide-treated fish indicated various physical and behavioral abnormalities including hypersecretion of mucus, erratic swimming, operculum movement, air gulping, tremors of fins, loss of equilibrium, and increased surface breathing. Histopathologically, gills tissues of treated fish indicated atrophied lamellae, uplifting of secondary lamellae, necrosis of primary and secondary lamellar epithelial cells, telogenesis, congestion, and lamellar fusion. Histopathological examination of liver tissues of treated fish showed mild to moderate congestion, necrosis of hepatocytes, and atrophy of hepatocytes while kidneys revealed degeneration of renal tubules, glomerular atrophy, ceroid, and necrosis of renal tubules. The erythrocyte counts, monocyte and lymphocyte counts, and hemoglobin values were significantly ( P < 0.05) reduced in pendimethalin-treated fish. Results on serum biochemistry showed that the biomarkers of kidneys, heart, and liver were significantly higher in fish of treated groups. In addition, values of different biochemical reactions like reactive oxygen species (ROS), thiobarbituric acid reactive species (TBARS), total proteins, and quantity of different antioxidant enzymes including reduced glutathione (GSH), catalase, and superoxide dismutase (SOD) were significantly different when compared to untreated fish. Moreover, the percentile of different nuclear abnormalities in red blood cells and frequency of DNA damage increased significantly in treated fish. It can be concluded from the findings that pendimethalin causes its toxic effects via disruption of physiological and hematobiochemical reactions of fish., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2022 Jin-qing Wang et al.)

Enterotoxemia is a severe and peracute disease caused by Clostridium perfringens ( C. perfringens ) rendering high mortality leading to huge economic losses, especially in small ruminants. The bacterium induces peracute death in animals based on the rapid production of different lethal toxins. Mortality occurred three private herds of two breeds, i.e., Makhi Cheeni and Beetal, and one non-descriptive (Teddy) herds reared in the desert area of Bahawalpur, Pakistan. At necropsy, tissue samples for histopathology and intestinal contents for bacterial isolation and culture were collected. Following the standard procedure, tissue slides were prepared. Multiplex PCR was used to identify toxinotypes using specific primers. Morbidity, mortality, and case fatality in Makhi Cheeni, Beetal, and Teddy goats caused by enterotoxemia were 87.58, 75.81, and 76.11%, respectively. Based on toxinotypes in the present outbreaks, C. perfringens type A ( cp α = 20.7%; cp α + cp β2 = 11.2%) and C. perfringens type D ( cp α + cp β2 + etx = 47.7%; cp α + etx = 20.7%) were detected. Deaths due to C. perfringens type D (68.10%) were significantly higher ( p < 0.001) compared with deaths by C. perfringens type A (34.90%). Petechiation of serosal surfaces, hemorrhage of intestines, lungs, and liver were seen. Kidneys were soft, and under the microscope, tubules were studded with erythrocytes. There was stunting and fusion in the intestinal villi. From this study, we concluded that endotoxemia can occur in any season; thus, a proper vaccination schedule must be followed for the protection of small ruminants' health., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hussain, Guangbin, Abbas, Siddique, Mohiuddin, Khan, Rehman and Khan.)

Mycotoxicosis is the second most important problem faced by the Pakistan poultry industry, after high feed prices. The present experimental study was designed to investigate the toxicopathological effects of aflatoxin B1 (AFB1) in commercial broiler chicks and its amelioration with locally produced mycotoxin binder. Total of 125 broiler chicks was divided into five equal groups (A-E). Group A served as negative control, group B (300 μg AFB1/kg feed) as positive control, group C (300 μg AFB1/kg + Local Mycotoxin Binder (LMB), 1 g/kg feed), group D (300 μg AFB1/kg + 2 g LMB/kg feed), and group E (300 μg AFB1/kg + Commercial Mycotoxin Binder (CMB), 2 g/kg of feed). Parameters studied included mortality, feed intake, bodyweights, absolute and relative organ weights, and gross and microscopic lesions in visceral organs. Clinical signs including alertness, fecal consistency, and feather shine were significantly lower in group B compared with control group A. The feed intake of 2 g/kg LMB treated group was significantly higher than that of the positive control group B. Also mean bodyweights of group D birds was higher than that of group B birds indicating an ameliorative effect of LMB. Histopathological results showed that moldy feed produced necrotic changes in the liver and kidneys in group B birds. However, in group D and E birds, the hepatic and renal parenchyma was normal, showing a protective effect of LMB. In the present study, a higher dose of LMB (2 g/kg) in group D showed higher bodyweights and feed intake. In group D, birds hepatic and renal parenchyma was also normal. The results suggested that local mycotoxin binder ameliorated the toxicopathological effects of AFB1 in mortality, feed intake, bodyweights, organ weights and, gross and microscopic lesions in visceral organs. These ameliorative effects of LMB were dose-dependent. The results of the present study concluded that AFB1 intoxication leads to decrease in bodyweights, feed intake in dose-related manner. The mortality was also dose-dependent. Gross and microscopic changes in the aflatoxin groups were more pronounced, however, all these deleterious effects were ameliorated in higher dose of LMB (group D) and CMB (group E). In group C, these deleterious effects were partially ameliorated. Local mycotoxin binder is an economical solution for aflatoxicosis problem, making poultry production more cost-effective., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Avian influenza virus subtype H9N2 is identified in chickens with respiratory disease while Bacillus cereus (B. cereus) has been frequently isolated from chicken feed in China. However, the roles of co-infection with these two pathogens remain unclear. In the present study, SPF chicks were intragastrically administered with 10 8  CFU/mL of B. cereus for 7 days and then inoculated intranasally with 100 EID 50 of H9N2 three days later. Alternatively, chickens were initially inoculated with H9N2 and then with B. cereus for one week. Post administration, typical respiratory distress persisted for 5 days in both co-infection groups. Gizzard erosions developed in the groups B. cereus/H9N2 and B. cereus group on 7 th day while in group H9N2/B. cereus on 14 th day. More importantly, both air-sac lesions and lung damage increased significantly in the co-infection group. Significant inflammatory changes were observed in the B. cereus group from day 7 to day 21. Moreover, higher loads of H9N2 virus were found in the co-infected groups than in the H9N2 group. Newcastle Disease Virus (NDV) specific antibodies were decreased significantly in the H9N2/B. cereus group compared to the B. cereus and the B. cereus/H9N2 groups. Nonspecific IgA titers were reduced significantly in the B. cereus group and the H9N2/B. cereus group compared to the control group. In addition to this, lower lymphocyte proliferation was found in the con-infection groups and the H9N2 group. Hence, feed-borne B. cereus contamination potentially exacerbates gizzard ulceration and aggravates H9N2-induced respiratory distress by inhibiting antibody-mediated immunity and pathogen clearance. Thus controlling the B. cereus contamination in poultry feed is immediately needed.

Demand for poultry meat is rising in low- and middle-countries, driving the expansion of large commercial farms where antimicrobials are used as surrogates for hygiene, good nutrition. This routine use of antimicrobials in animal production facilitates the emergence and spread of antibiotic-resistant pathogens. Despite potentially serious consequences for the animal industry, few studies have documented trends in antimicrobial use (AMU) at the farm-level in low- and middle-income countries. The objective of this study was to estimate AMU in a broiler chicken farm in Pakistan over a five-year period and to extrapolate national AMU in commercial broiler farming. Between 2013 and 2017, we monitored AMU in 30 flocks from a commercial broiler farm in Punjab, the most populous province of Pakistan. The amount of antimicrobials administered was calculated in milligram/population unit of the final flock weight (mg/fPU) and in used daily dose (UDD). The annual on-farm antimicrobial use was 250.84 mg of active ingredient per kilogram of the final flock weight. This consumption intensity exceeds the amount of antimicrobial used per kilogram of chicken of all countries in the world except China. Measured in mg per kg of final flock weight or population unit (fPU), medically important drugs such as colistin (31.39 mg/fPU), tylosin (41.71 mg/fPU), doxycycline (81.81 mg/fPU), and enrofloxacin (26.19 mg/fPU) were the most frequently used antimicrobials for prophylactic or therapeutic use. Lincomycin was the most frequently used antimicrobial used in-feed (29.09 mg/fPU). Our findings suggest that the annual consumption of antimicrobials in the broiler sector in Pakistan could be as high as 568 tons. This alarmingly high consumption estimate is the first baseline study on antimicrobial use in animals in Pakistan. Our findings call for immediate actions to reduce antimicrobial use in Pakistan, and countries with comparable farming practices.

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 (BCSP 31 ). For this purpose, the BCSP 31 gene was amplified, purified and ligated in pTZ57 R/T (cloning vector). Dubbed BCSP 31 -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-BCSP 31 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., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This study was conducted to investigate the toxic effects of fipronil on common carp (Cyprinus carpio). The trial was performed in laboratory of Department of Life Sciences at the Islamia University of Bahawalpur, Punjab, Pakistan. Total of 72 fishes, with apparently no clinical signs were purchased from local fish hatchery near Bahawalpur, Punjab, Pakistan. After 2 weeks of acclimatization fish were divided into six groups in glass aquaria (each having 100 liters of water capacity) and treated with different concentrations of fipronil for 12 days: 0 mg/L-control group (A),0.02 mg/L (B),0.04 mg/L (C),0.06 mg/L (D),0.08 mg/L (E) and 0.10 mg/L (F). Blood samples were taken every 4 days for evaluation of hemato-biochemical parameters. Fish in high doses treated groups show severe abnormalities in clinical-hematological and biochemical parameters. At high dose in group E and F fish show clinical ailments such as convulsions, jerking, faintness, increase in operculum movement, body curvature and breathing difficulty. Erythrocyte count, hemoglobin, hematocrit were decreased significantly (P < 0.05) and mean corpuscular volume, total leukocyte count, neutrophils, monocytes, and lymphocytes were significantly increased. Biochemical parameters including urea, creatinine, cholesterol, triglyceride, glucose were significantly increased but albumin was significantly decreased. Nuclear and cellular abnormalities were also increased in high dose treated groups. Hence fipronil induces clinico-hematological and serum biochemical changes in common carp C. carpio.

Hemoparasitic diseases like trypanosomiasis have an adverse influence on the health and working capability of infected animals. Monitoring and identification of blood born parasitic infections in dairy animals are of vital importance to get the optimum production. In this study blood samples were collected from Nili Ravi buffaloes (n = 390) kept at different villages of district Lodhran, Punjab province of Pakistan. Blood samples were evaluated for red blood cell counts, total and differential leukocyte counts, hematocrit, hemoglobin, total proteins and different serum parameters such as alanine aminotransferase, aspartate aminotransferase, malondialdehyde, alkaline phosphatase, lactate dehydrogenase, phosphorous, copper, calcium and magnesium. Overall prevalence of Trypanosoma evansi was 4.61% based on microscopic smear examination, 11.02% with Formol Gel Test and 16.15% with PCR. Infected buffaloes showed different clinical signs, including high fever (105 ± 1.0 °F), edema of face and legs, hyperemic mucosa of eyes, lachrymation, bulging eyes, pale mucus membranes and frequent urination. Microscopic examination of blood films showed morphologically different parasites. Statistical analysis did not indicate an association of infection based on age and sex of buffaloes. Results revealed significantly (p < 0.05) lower values of red blood cell counts, hemoglobin, hematocrit, mean corpuscular hemoglobin concentration and total proteins, while increased values of mean corpuscular volume, total white blood cells, monocyte, neutrophils, basophils and eosinophils in infected animals. Infected buffaloes were suffering from macrocytic hypochromic anemia. A significant (p < 0.05) increase in serum lipid per oxidation product (malondialdehyde) level and serum enzymes while a decrease in macrominerals and trace mineral (copper) in trypanosomiasis positive buffaloes were recorded. It was concluded that Trypanosoma evansi is prevalent in Pakistan under tropical and subtropical climatic conditions. It causes clinical disease with macrocytic hypochromic anemia and oxidative stress in infected buffaloes., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The current study was designed to investigate pathological effects of fowl adenovirus in broilers exposed to aflatoxin B 1 . 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 B 1 (AFB 1 ) 100 and 200 μg/kg feed. Group D was infected with FAdV-4, while groups E and F administered with both AFB 1 (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 + AFB 1 200 μg/kg feed) and the lowest mortality was observed in group B (AFB 1 100 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 AFB 1 intoxication lead to dose dependent changes in liver and kidneys. Severity of the changes was increased in interactive groups of AFB 1 with FAdV-4. Therefore, feed should be regularly monitored for AFB 1 levels and day old chicks for vertically transmitted FAdV-4 to prevent losses., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Peste des petits ruminants (PPR), an economically important viral transboundary disease of small ruminants is not only prevalent in Pakistan but also in other countries where people rely on agriculture and animal products. The present study was aimed at describing the pathology and antigen localization in natural PPR infections in local (Kajli sheep; Beetal goats) as well as imported small ruminant breeds (Dorper sheep; Australian Boer goat). Morbidity and mortality rates were significantly (P < 0.001) higher in indigenous Kajli sheep (75.37 and 32.80%) and Beetal goats (81.10 and 37.24%) as compared to Dorper sheep (6.99 and 1.48%) and Australian Boer goat (5.01 and 2.23%). Affected animals exhibited high fever, severe diarrhea, abdominal pain, respiratory distress and nodular lesions on lips and nostrils. Thick mucous discharge was oozing out from nostrils. On necropsy, lungs were congested and pneumonic, with nodular and cystic appearance. Intestines were hemorrhagic with zebra stripping. Characteristic histopathological lesions of PPR were noted in intestines, lymphoid organs and lungs. In GI tract, stunting and blunting of villi, necrotic enteritis, and infiltration of mononuclear cells in duodenum, jejunum and ileum. Small intestines exhibited diffuse edema of the submucosa along with proliferation of fibrocytes leading to thickened submucosa which has not been reported previously. Lymphoid organs showed partial to complete destruction of lymphoid follicles. Lesions of the respiratory tract included depictive of bronchopneumonia, severe congestion of trachea and apical lobe of lungs with deposition of fibrinous materials. Histopathological lesions of respiratory tract were severe and characteristic of broncho-interstitial pneumonia, bronchopneumonia, interstitial pneumonia and fibrinous pneumonia. The alveoli were filled with edematous fluid mixed with fibrinous exudate, numerous alveolar macrophages, mononuclear cells along with thickened interalveolar septa and presence of intranuclear eosinophilic inclusion bodies. One-Step RT-PCR using NP3 and NP4 primers confirmed a PPR virus of 352 bp size in spleen, lungs and mesenteric and brachial lymph node samples. It was concluded that morbidity and mortality due to PPR were significantly higher in indigenous breeds of sheep and goat as compared to imported sheep and goat breeds. PPR has rendered various lesions in GI and respiratory tract which are characteristic in nature for the diagnosis of the disease under field condition., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The present study was designed to investigate any ameliorative effects of bentonite (BN) against immuno-pathological alterations induced by dietary aflatoxin B 1 (AFB 1 ) or ochratoxin A (OTA) in broiler chicks. In one experiment, AFB 1 (0.1, 0.2 or 0.6 mg/kg feed) was fed alone and par alley with bentonite clay (3.7 or 7.5 g/kg feed) to the broilers. In the second experiment, the broilers were given feed contaminated with OTA (0.15, 0.3 or 1.0 mg/kg feed) alone and in combination with bentonite clay (3.7, 7.5, or 15 g/kg feed). Experimental feedings were continued for 42 days. At various time points along the feeding schedule, immune system organ histologic status, as well as host humoral and cellular immune responses, were evaluated in all groups. The dietary addition of AFB 1 and OTA alone significantly reduced immune responses in the birds as assessed by histological changes in the bursa of Fabricius and thymus, antibody responses to SRBC, in-vivo lympho-proliferative responses to Phytohemagglutinin-P (PHA-P) and, phagocytic function in situ. The dietary addition of BN significantly ameliorated the immunotoxicity of 0.1 and 0.2 mg/kg dietary AFB 1 , however with a level of 0.6 mg AFB 1 /kg only partial amelioration was seen. The co-treatment of birds exposed to OTA with BN at all levels only partially alleviated deleterious effects on histology and immune responses. Taken together, the results here suggested to us that dietary addition of BN could help ameliorate AFB 1 -mediated immunotoxicities but could not afford such protection against OTA-induced immune damage.

Since 2007, most areas of China have seen outbreaks of poultry airsacculitis, which causes hugely economic losses to the poultry industry. However, there are no effective measures to combat the problem. In this study, 105 rations were collected to isolate Aspergillus spp. from the diseased farms. In subsequent experiments, SPF chickens were inoculated with Ornithobacterium rhinotracheale (ORT), Chlamydia psittaci (C. psittaci) and Aspergillus fumigatus (A. fumigatus), and mortality rate, body weight gain and lesion score were evaluated. Of these ration samples, 63 (60.0%) were A. fumigates, 21 (20.0%) were Aspergillus niger (A. niger) and 11 (10.5%) were Aspergillus candidus (A. candidus). Furthermore, SPF birds infected with C. psittaci, ORT, H9N2 virus and A. fumigatus conidia exhibited a mortality rate of 40%, while simultaneous co-infection with C. psittaci, ORT and A. fumigatus resulted in a mortality rate of 20%. The avian airsacculitis was manifested in the C. psittaci + ORT/A. fumigatus, C. psittaci + H9N2 + ORT/A. fumigatus and C. psittaci + H9N2/A. fumigatus groups while others had transient respiratory diseases without mortality. Our survey indicates that feed-borne A. fumigatus is prevalent in poultry rations. The combination of C. psittaci, ORT, H9N2 and A. fumigatus conidia contributes to the replication of avian airsacculitis by aggravating the severe damage to the air sacs and lungs of chickens.

Mycoplasma bovis (M. bovis) is an important bacterium, causing severe respiratory infection, and arthritis in dairy animals worldwide. This study is based on 50 suckling calves among which 15 showed respiratory distress, lameness and swollen joints and died later. M. bovis was isolated and identified from all dead (n = 15) and live (17.14%; 06 out of 35) calves on the basis of bacteriological examination. In morbid calves, the carpus and stifle joints were severely affected, while necropsy revealed multiple well-circumscribed calcified abscesses and caseous exudates in cranio-ventral and diaphragmatic lobes of lungs. Suppurative polyarthritis, fibrino-suppurative, teno-synovitis and otitis media were the common and striking lesion observed at postmortem examination. Histopathological examination revealed broncho-interstitial pneumonia and necrotic fibrino-purulent broncho-pneumonia in lungs. Similarly, synovial membranes and joints revealed presence of multiple foci of liquefactive necrosis surrounded by lymphocytes, plasma cells, macrophages and peripheral fibroplasia. In the bacteriological investigations, the characteristic fried egg colonies of M. bovis further confirmed this infection in all suspected cases. In conclusion, the current clinico-histo-pathological features are the depictive picture, and is the first report of M. bovis infection in calves in Pakistan., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

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.

The present study aimed to investigate the deleterious effects of chlorpyrifos (CPF) in experimentally exposed broiler birds. The experiment was carried out on one day old (n = 120) broiler chicks. The CPF was reconstituted in corn oil as vehicle (1 ml/kg) to obtain a final concentration of a single dose to the birds 5, 10 and 20 mg/kg body weight (BW) for fourteen days of the experiment through the stomach tube. The control group was given corn oil 1 ml/kg only. Birds exposed to high dose (20 mg/kg BW) showed signs of toxicity (salivation, lacrimation, gasping, convulsions, frequent defecation and tremors). The birds exposed to 10 and 20 mg/kg showed significantly (P ≤ 0.05) decreased body weight. Significantly (P ≤ 0.05) decreased hematological parameters i.e. total erythrocyte counts, hemoglobin concentration, hematocrit and total leukocyte were observed in the high dosed group as compared to control and other low dosed fed birds. Serum protein and albumin showed a significant (P ≤ 0.05) increase in high dosed CPF fed birds. Non significant results were observed in the case of globulin. The acetylcholinestrease (AChE) activity was significantly (P ≤ 0.05) decreased in blood, serum and plasma in CPF fed birds compared to control birds. In CPF fed birds as compared to control birds we found significantly (P ≤ 0.05) higher levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Necrotic and degenerative changes were observed on histopathological investigations of spleen, kidneys, bursa of Fabricius, thymus and brain tissues in CPF exposed birds. In conclusion the chlorpyrifos induced toxicopathological effects on health biomarkers of broiler chicks., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The present study was carried to ascertain the association of various risk factors of mastitis in water buffaloes. The milk samples from buffaloes were collected and screened through California Mastitis Test for the presence of mastitis. In the present study, 15.2 % prevalence of subclinical mastitis was recorded both at the government (13.4 %) and private farms (15.5 %). The chi-square analysis showed significantly higher involvement of the right rear and front quarters. The analysis of variance technique showed significant difference in live body weight, milk yield, teat end to floor distance (P < 0.001), udder depth, teat length, and teat diameter in mastitic and healthy buffaloes. The frequency analysis also revealed significant difference between various groups including lactation stage, teat and/or udder pathology, teat shape, and udder shape (P < 0.001). The logistic regression analysis revealed significant positive association of mastitis with milk leakage, live body weight, milk yield, parity, calf suckling, pendulous udder, number of attendants at the farm, dirty hind legs, and udder depth.

This study aimed to evaluate the immunological status of progeny of hens kept on ochratoxin A (OTA)- and aflatoxin B(1) (AFB(1))-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 AFB(1)/Kg, while hens in Groups E and F were kept on feed amended with OTA and AFB(1) 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 + AFB(1) 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 AFB(1)-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 AFB(1) 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.

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., (Copyright © 2011 Society of Chemical Industry.)

The objective was to study the growth potential of Sahiwal calves given milk or milk replacer with or without concentrates. For this purpose, forty-eight Sahiwal calves were divided into four groups of 12 animals each with equal sex ratio. In each group, the calves were offered either milk or a milk replacer (MR) at a rate of 10% of their body weight adjusted weekly. In addition to this, calves were fed either a starter ration plus Egyptian clover hay (SR + H) or hay only (H) until the end of trial. The milk or MR was withdrawn gradually from day 56 until animals were weaned completely by day 84. Calves offered milk grew faster than those offered MR (357 ± 9 vs. 162 ± 9 g/day; p < 0.05) and displayed higher weaning weights (51.6 ± 0.8 vs. 35.2 ± 0.8 kg; p < 0.05). The calves offered SR + H grew faster (311 ± 9 vs. 208 ± 9 g/day; p < 0.05) and displayed higher weaning weights (48.7 ± 0.8 vs. 38.1 ± 0.8 kg; p < 0.05) than those fed H alone. Calves offered milk plus SR + H showed the highest growth rate and weaning weights (401 ± 13 g/day and 56.3 ± 1 kg, respectively). The lowest growth rate and weaning weights were observed in calves offered MR and H only (115 ± 13 g/day and 30.3 ± 1 kg, respectively). Calves offered the MR had higher number of scour days than those offered milk (13.5 vs. 3.3). The feeding of whole milk in combination with the starter ration and hay resulted in superior growth rates, higher weaning weights, and healthier calves than the other feeding regimens.

Seroprevalence, clinical findings, and lesions of contagious caprine pleuropneumonia (CCPP) in Beetal goats were recorded during an outbreak. The overall seroprevalence of CCPP was 32.50%. Confirmation of Mycoplasma mycoides in serum was carried out using counter immunoelectrophoresis (CIE) technique. The highest CIE-positive cases were recorded in the older goats (51.72%) as compared to young ones. Nasal swabs collected from 39 goats showing respiratory signs were found positive for M. mycoides. The most consistent clinical findings were mild to severe cough, purulent nasal secretion, emaciation, dyspnea, increased respiration rate, and pyrexia. Mortality due to CCPP was 9.17%. Consolidation of lungs exhibited the highest frequency (100%), followed by alveolar exudation (90.90%) and pleural adhesion (72.72%). Among the microscopic lesions, septal peribronchiolar fibrosis exhibited the highest frequency (81.81%), followed by fibrinous pleuritis (63.63%) and peribronchiolar cuffing of mononuclear cells (54.54%) in lungs. From these results, it was concluded that CCPP under subtropical conditions has high prevalence in Beetal goats and leads to significant mortality.

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 NaHCO(3) 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.

This study aimed to investigate the phagocytic potential of macrophages in progeny of breeder hens kept on an OTA-contaminated diet. For this purpose, 84 White Leghorn (WL) layer breeder hens (40-weeks-of-age) were divided into seven groups (A-G). Hens in Group A were fed a commercial layer ration while those in Groups B-G were kept on a diet amended with 0.1, 0.5, 1.0, 3.0, 5.0, or 10.0 mg OTA/kg, respectively, for up to 3 weeks (n = 12/treatment group; n = 4/time sub-group/treatment group). Fertile eggs were set for hatching on a weekly basis to get the progeny of each week separately. Hatched chicks (n = 10 from each group) were injected with India ink at day 14-of-age to study the in vivo phagocytosis of carbon particles. At day 30, abdominal macrophages were collected from 15 chicks/group and were used to assess their ex vivo/in vitro phagocytic potential against sheep red blood cells (SRBC) as well as for nitrite production upon challenge with lipopolysaccharide (LPS). The phagocytic indices of the reticuloendothelial system of all three sets of progeny (chicks obtained from hens fed OTA for 7, 14, and 21 days) were significantly lower than values seen with Group A chicks. The number of macrophages that were actively phagocytic, the number of SRBC internalized per macrophage, and the extent of nitrite production after stimulation with LPS were each significantly lower in the cells obtained from chicks of breeder hens that had been maintained on the OTA-contaminated diets. The findings of this study clearly showed that there are immunosuppressive effects-in terms of depressed in vivo and in vitro macrophage functionality-in progeny of OTA-fed breeder hens.

This study was planned and executed with the aims to explore corpus luteal primary cell culture as an "animal alternate testing system" in toxicity studies and in vitro toxic effects of cypermethrin (CY) 90% (pyrethroid) and methamidophos (MTP) 73% (organophosphate) on morphology and progesterone secretory activity of bovine corpus luteal cells and tissue. For this purpose, primary cell cultures of bovine corpus luteum (CL) cells were maintained in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 5% fetal calf serum (FCS). Toxicity evaluation were based on viable CL cell counts, morphological changes in CL cells, ability of CL cells to produce progesterone and histological changes in CL tissue at different hours post exposure to CY and MTP. The changes induced by both the insecticides were time and dose dependant. Viable cell counts and progesterone concentration decreased significantly (P<0.05) with the treatment of CY and MTP when compared to control. Corpus luteal cells exposed to CY showed more severe toxic effects as compared to MTP, though the difference was non-significant. Cellular or tissue alterations included degenerative changes in luteal cells, pleomorphic changes, nuclear degeneration and vacuolation, cell shrinkage and rupture, cloudy swelling and hydropic degeneration, less cytoplasmic granulation, cell elongation, hyalinization and cytoplasmic haziness and stripling and necrosis. It was concluded that both the insecticides induce toxic effects in terms of viable counts, morphological and histological changes and progesterone production of bovine CL cells. Cypermethrin exhibited more adverse toxic changes in viable cell counts, progesterone production and histological findings as compared to methamidophos., (Copyright © 2009 Elsevier GmbH. All rights reserved.)

The objective of the present study was to determine the pathological and genotoxic effects of atrazine (ATZ) in male Japanese quail. Adult male Japanese quail were administered ATZ daily at 0-500 mg/kg bw (A-H groups) orally for 45 days. The blood and morbid tissues were collected at day 15, 30, and 45 of the treatment. A significant decrease in feed intake, body weight, erythrocyte counts, hemoglobin and hematocrit values were observed at high ATZ dose compared to control. Leukocyte counts decreased significantly throughout the experiment in groups E-H (50-500 mg/kg bw). Grossly, testes from ATZ treated birds were comparatively smaller in size. Histologically, seminiferous tubules of testes in group H (500 mg/kg bw) exhibited decreased number of spermatocytes, necrotic nuclei of spermatids, and lesser number or absence of spermatozoa. Biliary hyperplasia and vacuolar degeneration in liver and mild renal tubular necrosis was observed in birds higher doses. Significantly longer comet tails of DNA damage in leukocytes and isolated hepatocytes were recorded with 500 mg/kg bw ATZ.

Fifty female Japanese quail (Coturnix japonica) at 40 days were divided into five equal groups. Sodium hypochlorite was added in drinking water to provide 0, 25, 50, 200, 400 mg chlorine/L for 6 weeks. At week 7, the chlorine concentration of 25 and 50 mg groups was increased to 100 mg and further increased to 400, 1600 and 6400 mg at weeks 8, 9 and 10, respectively. At the end the experiment (week 10), all the birds were sacrificed. No clinical signs were observed in quail given 50 mg chlorine/L. Quail given 200-400 mg/L chlorine exhibited decreased feed intake, body weight and egg production. In addition, those given 1600-6400 mg/L chlorine also exhibited depression. Decreased hematocrit, erythrocyte, hemoglobin, leukocytes, plasma proteins and globulin were observed at 200 mg/L chlorine and higher concentration. Gross lesions were not observed in quail offered 200-400 mg/L chlorine for 6 weeks; however, those offered 1600-6400 mg/L chlorine exhibited atrophied pectoral muscles, prominently protruding keel bones, decreased weight of ovary and oviduct and a reduction in an area of the different segments of oviduct. The histopathological picture in mucosa of oviduct consisted of degenerated glandular cells. In some cases, glandular tissue was replaced by cord of cells and fibroblast. The present study suggested that sodium hypochlorite in drinking water yielding 50 mg/L chlorine was not toxic for 6 weeks. However, sodium hypochlorite providing 200 mg/L chlorine or higher concentrations in drinking water was toxic to the quail.

The objective of the present study was to explore modification in toxico-pathological responses of rats toward aflatoxins in the presence of cypermethrin. A total of 120 adult male rats divided into six equal groups received AF and cypermethrin alone or in different combinations. AF was administered daily into rats with a stomach tube at dose rates of 0, 0.5 and 1.0mg/kg AFB1. Cypermthrin was administered in the feed at dose levels of 0 and 500mg/kg. Rats administered AF alone showed depression, decrease in feed intake, body weight and loose feces. Livers exhibited fatty change, necrosis, newly formed bile ducts and increased diameter of hepatocytes and their nuclei. Lesions in kidney included tubular necrosis and pink homogeneous tubular casts. Serum ALT and creatinine concentrations increased while those of total proteins, albumin, serum cholesterol and triglycerides decreased. Rats fed cypermethrin only had decreased feed intake and body weight. Hepatocytes showed fatty change and cellular necrosis. A concurrent administration of AF with cypermethrin indicated a potentiation of the AF toxicity reflected by increased severity of clinical signs, mortality of the rats and decreased body weights. Kidneys' relative weight also showed an equivocal interaction between the two toxicants. Other parameters studied did not show significant differences between the rats administered AF alone or concurrently with cypermethrin.

Background: Degenerative suspensory ligament desmitis (DSLD) is a debilitating disorder thought to be limited to suspensory ligaments of Peruvian Pasos, Peruvian Paso crosses, Arabians, American Saddlebreds, American Quarter Horses, Thoroughbreds, and some European breeds. It frequently leads to persistent, incurable lameness and need to euthanize affected horses. The pathogenesis remains unclear, though the disease appears to run in families. Treatment and prevention are empirical and supportive, and not effective in halting the progression of the disease. Presently, the presumptive diagnosis of DSLD is obtained from patient signalment and history, clinical examination, and ultrasonographic examination of clinically affected horses, and is confirmed at post mortem examination. Presently, there are no reliable methods of diagnosing DSLD in asymptomatic horses. The goal of this study was to characterize and define the disorder in terms of tissue involvement at the macroscopic and microscopic levels., Results: We examined tissues and organs from 28 affected horses (22 Peruvian Pasos, 6 horses of other breeds) and from 8 control horses. Histopathological examination revealed the presence of excessive amounts of proteoglycans in the following tissues removed from DSLD-affected horses: suspensory ligaments, superficial and deep digital flexor tendons, patellar and nuchal ligaments, cardiovascular system, and sclerae. Electron microscopy demonstrated changes in diameters of collagen fibrils in the tendon, and in smooth muscle cells of the media of the aorta compatible with increased cell permeability in DSLD-affected cells. Separation of tendon extracts by gel chromatography revealed the presence of additional proteoglycan(s) in extracts from affected, but not control extracts., Conclusion: This study demonstrates for the first time that DSLD, a disease process previously thought to be limited to the suspensory ligaments of the distal limbs of affected horses, is in fact a systemic disorder involving tissues and organs with significant connective tissue component. Abnormal accumulation of proteoglycans between collagen and elastic fibers rather than specific collagen fibril abnormalities is the most prominent histological feature of DSLD. Because of this observation and because of the involvement of many other tendons and ligaments beside the suspensory ligament, and of non-ligamentous tissue we, therefore, propose that equine systemic proteoglycan accumulation or ESPA rather than DSLD is a more appropriate name for this condition.