In vivo analysis the effect of antibiotic growth promoters (AGPs), Oxytetracycline di-hydrate and Tylosin phosphate on the intestinal microflora in broiler chicken.

The study was aimed to analyse the effects of antibiotic growth promoters (AGPs), Oxytetracycline di-hydrate and Tylosin phosphate on the intestinal microflora in broiler chicken. The AGPs were provided in different concentrations solely or in combinations for 42 days of rearing. Faecal samples were collected from the intestine (duodenum, jejunum and caeca) of broiler chicken on 14th, 28th and 42nd days of trial. Samples were cultured on different selective medium and bacterial identification was performed by different biochemical and molecular diagnostic tools. Results showed a significant effect of AGPs on the growth of pathogenic microorganisms such as Escherichia coli and Clostridium perfringens in the intestine. Interestingly, an impaired growth was observed for both bacterium showing a significant effect (P<0.05) of AGPs on E. coli and C. perfringens on day 14th, 28th, and 42nd. This effect was observed solely and in combination while using AGPs. Data further showed that the effect was more prominent in combination and with an increase concentration of AGPs. Remarkably, no impairment was seen on the growth of L. reuteri at different sites of intestine and duration (14th, 28th, and 42nd days). The results showed that the use of AGPs in diet has no harmful effect on beneficial bacteria, however, an impaired growth was seen on the harmful bacteria. It is suggested that a combination of AGPs (OXY-1.0+TP-0.5) is economical and have no harmful effect on the broiler chicken. The use of AGPs in a recommended dose and for a specific period of time are safe to use in poultry both as growth promoter and for the prevention of diseases.

Molecular epidemiology, associated risk factors, and phylogenetic analysis of anaplasmosis in camel.

Camel Anaplasmosis is caused by members of family Anaplasmatacae, a tick transmitted, obligate intracellular bacteria. The etiological bacteria are transmitted by ixodid tick species. The species have multi host range distribution that is why it is crucial to diagnose it timely. The aim of present study was to investigate the molecular epidemiology i.e. prevalence and risk factors analysis of camel anaplasmosis. Furthermore, variations in hematological standards were also evaluated. The study found an overall 13.33% prevalence in camels. The confirmation of PCR positive samples for Anaplasma spp. was made through sequencing, the study isolatesshowed high homology with Iranian, Chinese, Philippines and South African isolates of Anaplasmatacae (Accession numbers'; KX765882, KP062964, KY242456, LC007100 and U54806) on BLAST queries. The phylogenetic analysis revealedthree study isolates of present study clustered with each other and the cluster was found closer to Chinese isolate of A. phagocytophilum (KY242456), A. marginale (KU586048), and Mongolian isolates of A. ovis (LC194134). Two of the isolates resembled Iranian isolate of Candidatus Anaplasmacamelii (KX765882), while one isolate resembled with Chinese isolates of A. Platys (KX987336) and Croatian isolates of A. Platys (KY114935). The key risk factors odds ratio (OR>1) identified for occurrence of camel anaplasmosis using regression model found sex and age of animal, previous tick history, tick infestation and tick control status, housing type, cracks in walls, rearing system and other species in surrounding as the key risk factors. The hematological parameters like lymphocytes, monocytes, granulocytes and platelets count were significantly decreased (p < 0.05) in diseased camels than healthy. This is the first ever molecular data on camel anaplasmosis in Pakistan. The disease should be monitored unceasingly as the etiologies have multi host distribution. Prompt attention should be offered to animals because neutropenia, lymphopenia and thrombocytopenia can exacerbate the disease by making the animal predisposed to otherdiseases.

Tetramethylpyrazine inhibited hypoxia-induced expression of calcium-sensing receptors in pulmonary artery smooth muscle cells in chickens.

Tetramethylpyrazine (TMP) is a biologically active ingredient, which is isolated from a popularChinese medicinal plant. It has been used effectively to treat ischemic heart problems, cerebrovascular and thrombotic vascular diseases. This study was designed to evaluate the effect of TMP on calciumsensing receptors in pulmonary artery smooth muscle in chickens. For this purpose forty day-old chicks were distributed into five groups: the control group, the hypoxia group (kept under low Oxygen treatment), and TMP groups (kept under low Oxygen treatment along with treatment of different concentrations of TMP). The pulmonary artery smooth muscle cells were also cultured on 6-well plates in high glucose culture medium and divided into the same five groups. We used in vivo and in vitro study models by applying immunohistochemistry, RT-qPCR assay and Western blotting analysis. Our results showed that pre-incubation with hypoxia markedly stimulated the activation of calcium-sensing receptor (CaSR) in pulmonary artery smooth muscle cells (PASMCs). The TMP decreased the mRNA and protein levels of CaSR. Treatment with TMP clearly inhibited the activation of all CaSR in a dose-dependent manner. Our data demonstrated that TMP can down-regulate the expression of CaSR. Therefore, these findings provide a new target to treat pulmonary arterial hypertension (PAH) under hypoxic conditions.

Protective effects of Herpetospermum caudigerum extracts against liver injury induced by carbon tetrachloride in mouse.

Herpetospermum caudigerum (H. caudigerum; HC), popularly known as “Sejimeiduo” in Tibet, it is widely used in Tibetan traditional medicine for the treatment of dyspepsia, liver and colic diseases. This study was designed to evaluate the effect of H. caudigerum extract (HCE) on suppressing liver injury induced by carbon tetra chloride (CCl4). For this purpose, we used CCl4 to induce acute liver injury in mouse model. The protective effects of HCE against liver injury were evaluated by biochemical parameters, histopathological and immunohistochemical staining. The results showed that the superoxide dismutase (SOD) activity was significantly increased with the increasing dose of HCE as compared to the CCl4-treated group (p less than 0.01); while AST and ALT levels in serum, MDA and MPO in liver were reduced in a dose-dependent manner. The histopathology showed that HCE treatment promoted the recovery of histopathological changes in liver in a dose-dependent way. Meanwhile, there was a higher expression of caspase-3 and NF-κB in the nucleus of several liver cells in the CCl4-induced group, and a low expression of caspase-3 and NF-κB were found with the increasing dose of HCE. Therefore, the present study suggests that HCE is a potent hepatoprotective agent that can treat acute liver injury and this ability may be attributed towards its anti-inflammatory and antioxidant potential.

Effect of CuSO4 and nano copper on serum antioxidant capacity in Weaned piglets.

Copper is the most essential trace element in the animal body. Nano-sized copper particles have been widely used in a number of different animal species in modern medicinal practice. The present study was designed to examine the effect of dietary copper sulfate/(CuSO4) and nano copper supplementation on serum antioxidant capacity of weaning piglets. A total of 28 Duroc piglets (21 days, and weighing ~7 kg) were randomly divided into three equal groups. The control group (n=4) was administered with a normal standard diet; however the CuSO4 (n=12) and nano copper (n=12) groups were treated with 50, 100 and 200 mg/kg/day body weight, respectively. After 28 days, blood serum copper-zinc superoxide dismutase (CuZn-SOD), ceruloplasmin (CP), malondialdehyde (MDA), myeloperoxidase (MPO), total antioxidant capacity (T-AOC), peroxidase (POD), nitric oxide (NO), nitric oxide synthase (NOS), hydrogen peroxide (H2O2) and inhibition of hydroxyl radical (CIHR) were analyzed from all groups. The results indicated that nano copper supplementation has significant (P less than 0.05) effect on the serum antioxidant capability as compared to dietary CuSO4 group in weaned piglets. Nano-size copper 100 mg/kg/day supplementation was confirmed to improve the immunity level by strengthening the antioxidant capacity of weaning piglets. Dietary supplementation with 100 mg/kg body weight nano copper could be a potential substitute for weaned piglets.

Correlation analysis of CD4 gene polymorphism and blood routine Indexes in pigs (Sus scrofa).

Cluster of differentiation 4 (CD4) is an important molecule in the immune system of animals, which participates in the processes of T-lymphocyte differentiation, maturation, immune response and signal transduction. During this study, we adopted the direct sequencing of PCR products and time-of-flight mass spectrometry methods for the detection of single nucleotide polymorphisms (SNPs) in 382 Yorkshire pigs. The linkage disequilibrium of CD4 gene polymorphisms and their genetic effects on blood routine indexes were also analyzed. The results showed that SNP was screened in intron 1, intron 6 and exon 10, respectively, of the porcine CD4 gene, and each SNP locus was detected in Yorkshire pigs, which had three genotypes with moderate polymorphism. The three SNPs were in strong linkage disequilibrium (r2>0.8), and constituted only two major haplotypes, AGT and CAC (both frequencies accounted for about 97%). The association analysis showed that CD4 gene polymorphisms were significantly correlated with white blood cell, lymphocyte and monocyte count in Yorkshire pigs (P less than 0.05). The white blood cell count and monocyte count in individual animals with CAC/CAC diplotype were significantly higher than those with AGT/CAC and AGT/AGT diplotypes (P less than 0.05). The lymphocyte count in animals with CAC/CAC diplotype were significantly higher than those with the AGT/AGT diplotype (P less than 0.05). This study indicates that CD4 gene is significantly associated with partial blood routine indexes in pigs, and it can be considered as a candidate gene for the study of porcine disease resistance.

Protective effects of Nano-elemental selenium against chromium-vi-induced oxidative stress in broiler liver.

The valuable role of selenium in mitigation of oxidative stress and heavy metal toxicity is well-known. Thus, the aim of the current study on broiler chickens was to examine whether nano elemental selenium (Nano-Se) supplementation can reduce the effects of chromium VI (K2Cr2O7) toxicity. For this purpose, a total of 150, one-day-old broiler chickens were allotted to five groups with three replicates: control group (standard diet), poisoned group (K2Cr2O7 via drinking water), protection group (K2Cr2O7 + Nano- Se), cure group (K2Cr2O7 for initial 2 weeks and then Nano-Se), and prevention group (opposite to the cure group). The broilers were detected by the activities of marker enzymes and oxidative stress markers including, aspartate aminotransferase (AST), alanine transaminase (ALT), gamma-glutamyl transferase (GGT) and superoxide dismutase (SOD), glutathione peroxidase (GSH-px), malondialdehyde (MDA), respectively. The (K2Cr2O7 administration caused histopathological damage in the liver of the chickens. Moreover, changes in serum biochemical indicators and oxidative stress parameters were also observed. Nano-Se supplementation increased the levels of GSH-px but reduced the activities of SOD, MDA, GGT, ALT and AST in the experimental groups (P less than 0.05). Our results showed that Nano-Se plays a protective role by preventing the oxidative stress induced by the chromium VI in broiler chickens.

FK228 recovers thiram-induced tibial dyschondroplasia in chicken via hypoxia inducible factor-1α.

Tibial dyschondroplasia (TD) is a disease of many avian species characterized by an enlarged and avascular lesion in the proximal tibiotarsal bone. The aim of present study was to evaluate the effects of hypoxia-inducible factor-1α(HIF-1α) inhibition on thiram- induced TD using synthetic medicine FK228 and the association between HIF-1α and heat-shock protein 90 (Hsp90). One hundred and fifty broiler chicks were equally divided into 3 groups: control; thiram fed; and FK228 treatment. Expressions of HIF-1α and Hsp90 genes were analyzed by real-time quantitative polymerase chain reaction (RT-qPCR) on day 10 and 14 post-hatch. Western blot analysis of HIF-1α and Hsp90 gene was performed to measure the protein levels at the end of the experiment. Results showed that HIF-1α and Hsp90 levels were significantly (P less than 0.05) up-regulated in the thiram group as compared to the control group. Meanwhile, FK228 (HIF-1α inhibitor) significantly (P less than 0.05) down- regulated the mRNA and protein levels of HIF-1α and Hsp90, restored the size of growth plate and diminished lameness. In conclusion, HIF-1α and Hsp90 play an important role in the formation of avascular growth plate and there is a direct relationship between HIF-1α and Hsp90 for the progression of TD pathogenesis. Therefore, HIF- 1α may prevent and control TD in broiler chickens.

Epidemiological investigation and risk factors of Peste des petitis ruminants (PPR) in yaks (Bos grunniens) and cattle in five regions of China.

To investigate the prevalence of Peste des petitis ruminants in yaks and cattle in poorly studied areas of China. A total of 1202 and 560 blood samples were collected from yaks and cattle, respectively and processed using a commercial ELISA kit. Multivariable logistic regression model was piloted to find the variables, potentially associated with exposure of PPR infection in yaks and cattle. Results indicated that the overall prevalence of PPR in yaks was 11.2%. A total 66 (11.8%) out of 560 were examined out to be positive to PPR in cattle in Jiangxi province. According to conditional stepwise logistic regression, region, age and gender were found to be the more influencing risk factors in yaks, while region and age were found to be the potential risk factors in cattle. The current results reported the prevalence and associated risk factors of PPR in bovine for the first time in China.

Antibiotic resistance and screening of the resistant genes of Escherichia coli (E. coli) isolated from diarrheal yak calves in Sichuan Province, China.

This study was conducted to determine the antibiotic and screening resistance genes of Escherichia coli (E. coli) isolated from diarrheal yak calves from high remote plateau in Sichuan, China. A total 41 rectal swabs were obtained from diarrheal yak calves. E. coli were isolated and identified. The antimicrobial sensitivity was tested by piloting the disk diffusion method for 21 antibiotics. Polymerase chain reaction was employed to detect the resistance genes. The results showed that the drug resistance ranged from 2.4% (amikacin) to 53.7% (tetracycline), while no isolates were found resistant to neomycin and polymyxin B. Multi-drug resistance was detected in 4.9% isolates to 17 antimicrobial agents; and 24.4% isolates were found susceptible to all antimicrobial agents. The aminoglycoside resistance genes of aac(3)-lla, ant(3')-la and aph(3')-lla was positive in 4.9%, 2.1% and 7.3% E. coli isolates respectively. The 4.9% and 2.1% of E. coli isolates were detected in b-Lactam resistance genes of TEM and CTX-M, respectively; and 12.2% and 4.9% of E. coli isolates were found to have Tetracycline resistance genes of tetM and tetA, respectively. The present study reveals that the yak calves from high cold plateau are potential reservoir of E. coli with widely distributed multiple drug resistance which requires the attention of concerned authorities regarding the use of non-standard antibiotics.

Epidemiological investigation and risk factors of Peste des petitis ruminants (PPR) in yaks (Bos grunniens) and cattle in five regions of China

@#To investigate the prevalence of Peste des petitis ruminants in yaks and cattle in poorly studied areas of China. A total of 1202 and 560 blood samples were collected from yaks and cattle, respectively and processed using a commercial ELISA kit. Multivariable logistic regression model was piloted to find the variables, potentially associated with exposure of PPR infection in yaks and cattle. Results indicated that the overall prevalence of PPR in yaks was 11.2%. A total 66 (11.8%) out of 560 were examined out to be positive to PPR in cattle in Jiangxi province. According to conditional stepwise logistic regression, region, age and gender were found to be the more influencing risk factors in yaks, while region and age were found to be the potential risk factors in cattle. The current results reported the prevalence and associated risk factors of PPR in bovine for the first time in China.

Transperineal template-guided prostate saturation biopsies in men with suspicion of prostate cancer: a pilot study from Pakistan.

Traditionally, transrectal ultrasound (TRUS)-guided biopsies are done for the diagnosis of prostate cancer (PCa) in Pakistan. The transperineal template-guided saturation biopsy (TTSB) approach has been recently introduced in Pakistan and we share diagnostic yields and pathological findings of specimens taken for PCa diagnosis in men with elevated serum total prostate specific antigen (PSA) and negative TRUS-guided prostate biopsies. In all, 16 patients investigated at the Department of Urology, Sindh Institute of Urology and Transplantation (SIUT), underwent TTSB. The mean age of patients was 67.8 ± 8.8 (range: 55 - 84) years. The median PSA was 9.5 (IQR: 7.9 - 19.8) ng/ ml. The duration of symptoms before biopsy ranged from 1 month to 144 months. The prostate was enlarged with mean weight of 73.5 ± 55.5 g. Histopathology revealed PCa in 5 of 16 (31.2%) cases. The Gleason score was 6 (3+3), 7 (3+4) and 8 (4+4) in 1 case each (6.3%) and 10 (5+5) in 2 cases (12.5%). At least two cores were positive in all positive cases. None of the patients required antibiotics post-procedure. In conclusion, the TTSB technique is a promising option for patients with elevated PSA level and negative transrectal prostate biopsies for the detection of PCa in our setting.

Prevalence, associated risk factors, and phylogenetic analysis of Echinococcus granulosus isolated from freerange Tibetan pigs in Tibet, China.

The current study was performed to investigate the prevalence, associated risk factors exploration and phylogenetic analysis of Echinococcus granulosus (E. granulosus) genotypes isolated from Tibetan pigs. A total 373 Tibetan pigs were examined during 2014 and 2015, and the variables potentially associated with E. granulosus infection were explored with a multivariable logistic regression model. E. granulosus cysts (n=37) were collected from Tibetan pigs (lungs or livers). Fragments amplification of mitochondrial (mt) DNA of cox1 (shorter and longer) and atp6 were employed. The genotype of E. granulosus were identified by sequence and phylogenetic analysis. Results showed the prevalence of E. granulosus in Tibetan pigs was 9.9%. The prevalence of E. granulosus in male and female Tibetan pigs was 6.8% and 13.3%, with a significant difference in the two genders (P<0.05). In different seasons, the infection rate of E. granulosus in Tibetan pigs were ranged from 5.8% to 12.3%. E. granulosus infection rates in different growing stages of Tibetan pigs were ranged from 4.4% to 15.9%, with a statistical difference in the three stages (P<0.01). The prevalence of E. granulosus in Tibetan pigs were 7.9% in 2014 and 13.0% in 2015. Genders and growing stages were demonstrated to be risk factors to influence the prevalence significantly through multivariable logistic regression model. All the three fragments were successfully amplified from each of the 37 cysts. E. granulosus genotypes of G4 and G6 were identified by comparing with reference sequences of E. genotypes available at NCBI database and phylogenetic analysis by using MEGA software.

Health status and renal function evaluation of kidney vendors: a report from Pakistan.

Unrelated kidney transplants have lead to commerce and kidney vending in Pakistan. This study on 104 vendors reports demographics, history, physical and systemic examination, ultrasound findings, renal and liver function and GFR by Cockcroft-Gault. Results were compared with 184 age, sex and nephrectomy duration matched living-related donors controls. Comparison of vendors versus controls showed mean age of 30.55 +/- 8.1 versus 30.65 +/- 7.85 (p = 0.91) years, M:F of 4.5:1 versus 4.2:1 and nephrectomy period of 33.89 +/- 30 versus 32.01 +/- 29.71 (p = 0.60) months respectively. Of the vendors 67% were bonded laborers earning <50 $/month as compared to controls where 68% were skilled laborers and self-employed earning >100 $/month. History of vendors revealed jaundice in 8%, stone disease in 2% and urinary tract symptoms in 4.8%. Postnephrectomy findings between vendors versus donors showed BMI of 21.02 +/- 2.8 versus 23.02 +/- 4.2 (p = 0.0001), hypertension in 17% versus 9.2% (p = 0.04), serum creatinine (mg/dL) of 1.17+/-0.21 versus 1.02 +/- 0.27 (p = 0.0001), GFR (mL/min) of 70.94 +/- 14.2 versus 95.4 +/- 20.44 (p = 0.0001), urine protein/creatinine of 0.150 +/- 0.109 versus 0.10 +/- 0.10 (p = 0.0001), hepatitis C positivity in 27% versus 1.0% (p = 0.0001) and hepatitis B positive 5.7% versus 0.5% (p = 0.04), respectively. In conclusion, vendors had compromised renal function suggesting inferior selection and high risk for developing chronic kidney disease in long term.

Seroprevalence of HBV, HCV and HIV infections among college going first time voluntary blood donors.

OBJECTIVE: College going first time voluntary blood donors were tested for HBV, HCV and HIV infections, to know the seroprevalence of these diseases among them. METHOD: Blood donation camps were organized at 18 colleges and universities of the city and a total of 612 samples from first time donors were tested for HbsAg, anti-HCV, anti-HIV1, 2 antibodies on Abbott Diagnostic systems. RESULTS: Thirteen voluntary blood donors (2.21% with 95% confidence limit 1.2-3.52%) were HbsAg and 3 (0.5% with 95% confidence limit 0.12-1.33%) were anti HCV positive. None of the donors was positive for HIV infection. CONCLUSION: Seroprevalence of HBV, HCV and HIV infections among college going students is significantly low (< 3.0%) than 30% seroprevalence among paid donors and 7% among family/replacement blood donors. There is a need to educate, motivate and recruit college going students to regularly donate blood to ensure relatively safe supply of blood for transfusion services.