Khalid Mehmood

Antibiotic resistance, serogroups, virulence genes, and phylogenetic groups of Escherichia coli isolated from yaks with diarrhea in Qinghai Plateau, China.

BackgroundRuminants serve as one of the most important reservoirs for pathogenic Escherichia coli. Infection with E. coli, a foodborne enteropathogen, can lead to asymptomatic infections that can cause life-threatening complications in humans. Therefore, from a clinical and human health perspective, it is important to know which virulence genes, phylogenetic groups, serogroups, and antibiotic resistance patterns are present in E. coli strains in yaks with diarrheic infections.MethodsTwo-hundred and ninety-two rectal swabs were collected from diarrheic yaks in Qinghai Plateau, China. The antimicrobial sensitivity of each resulting isolate was evaluated according to the disk diffusion method, and different PCR assays were performed for the detection of virulence genes and different phylogroups. Additionally, strains were allocated to different serogroups based on the presence of O antigen via the slide agglutination method.ResultsAmong the E. coli isolates tested, most of the isolates were multidrug resistant (97%) and harbored at least one virulence gene (100%). We observed ten virulence genes (sfa, eaeA, cnf1, etrA, papC, hlyA, aer, faeG, rfc, and sepA), of which sfa was the most commonly found (96.9%). Significant positive associations between some resistance phenotypes and virulence genes were observed (Pxa0<xa00.05, ORxa0>xa01). The majority of the E. coli isolates belonged to phylogroup A (79.5%), and the others belonged to phylogroups B1 (7.5%), D (4.1%), B2 (5.8%), and F (0.7%). Among all the E. coli strains tested, serogroups O91 and O145 were the most prevalent, accounting for 15.4 and 14.4%, respectively.ConclusionsOur results suggest that yaks with diarrhea serve as a reservoir of pathogenic E. coli carrying various virulence genes and resistance phenotypes. Therefore, clinicians and relevant authorities must ensure the regulatory use of antimicrobial agents and prevent the spread of these organisms through manure to farm workers and food-processing plants.

Intestinal epithelial cell injury induced by copper containing nanoparticles in piglets

The nano copper has been widely used in modern clinical medicine practice. However, it has been noticed that nano copper particles induce cell injury and toxicity. The present study was designed to determine the effect of nano copper particles on cell injury of intestinal epithelial cells (IECs) in piglets. The IECs were treated with different doses of nano copper (5, 10, 20 and 40μg/ml) for 24-48h to observe cell injury and toxicity. Cell injury was measured based on morphological and other changes including oxidative stress and genes expression. The oxidative stress markers were assayed by differential expression levels of SOD, MDA and Metallothionein (MT) in addition to CTR1, SOD1, COX17, MT and ATOX1 genes expression. Cellular morphology showed an increasing growth of cells without nano copper treatment and nano copper showed significant damage to IECs with higher dose as compared to low dose. Higher doses of copper nanoparticles (10, 20 and 40μg/ml) have membrane damaging effect on the intestinal epithelial cells, whereas MDA contents and MT value were significantly increased, and SOD activity was decreased with the increase in concentration of nanoparticles. Nano copper up-regulated the CTR1 and SOD1 genes and down-regulated the relative expression of COX17, MT and ATOX1 genes significantly in a dose-dependent manner. The findings of the current study provide important insights that nano copper plays an important role in intestinal epithelial cell injury and oxidative stress.

Icariin Ameliorate Thiram-Induced Tibial Dyschondroplasia via Regulation of WNT4 and VEGF Expression in Broiler Chickens

Tibial dyschondroplasia (TD) is main bone problem in fast growing poultry birds that effect proximal growth plate (GP) of tibia bone. TD is broadly defined as non-vascularized and non-mineralized, and enlarged GP with tibia bone deformation and lameness. Icariin (Epimedium sagittatum) is a traditional Chinese medicine, which is commonly practiced in the treatment of various bone diseases. Recently, many researcher reports about the beneficial effects of icariin in relation to various types of bone conditions but no report is available about promoting effect of icariin against TD. Therefore, current study was conducted to explore the ameliorating effect of icariin in thiram-induced TD chickens. A total of 180 broiler chicks were equally distributed in three groups; control, TD induced by thiram (50 mg/kg), and icariin group (treated with icariin @10 mg/kg). All groups were administered with normal standard diet ad libitum regularly until the end of experiment. The wingless-type member 4 (WNT4) and vascular endothelial growth factor (VEGF) genes and proteins expression were analyzed by quantitative real-time polymerase chain reaction and western blot analysis respectively. Tibial bone parameters, physiological changes in serum, antioxidant enzymes, and chicken growth performance were determined to assess advantage and protective effect of the medicine in broiler chicken. The expression of WNT4 was decreased while VEGF increased significantly (P < 0.05) in TD affected chicks. TD enhanced the GP, lameness, and irregular chondrocytes, while reduced the liver function, antioxidant enzymes in liver, and performance of chickens. Icariin treatment up-regulated WNT4 and down-regulated VEGF gene and protein expressions significantly (P < 0.05), restored the GP width, increased growth performance, corrected liver functions and antioxidant enzymes levels in liver, and mitigated the lameness in broiler chickens. In conclusion, icariin administration recovered GP size, normalized performance and prevented lameness significantly. Therefore, icariin treatments are encouraged to reduce the incidence of TD in broiler chickens.

Tibial dyschondroplasia is closely related to suppression of expression of hypoxia-inducible factors 1α, 2α, and 3α in chickens

Tibial dyschondroplasia (TD) cases has not been reported in Tibetan chickens (TBCs), but it is commonly seen in commercial broilers characterized by lameness. The underlying mechanism remains unclear. Hypoxia-inducible factors (HIFs) are important regulators of cellular adaptation to hypoxic conditions. In this study, we investigated the role of HIF-1α, -2α, and -3α in hypoxia and thiram-induced TD and their effect on tibial growth plate development in Arbor Acres chickens (AACs) and TBCs. RNA and protein expression levels of HIF-1α, -2α, and -3α were determined by using quantitative reverse transcriptase polymerase chain reaction and western blotting analyses, respectively. Interestingly, the results showed that HIF-1α, -2α, and -3α expressions in the tibial growth plate of TBCs were upregulated by hypoxia and the change was more significant in TBCs than in AACs. However, these factors were downregulated in thiram-induced TD. To further clarify the effect of thiram on tibial growth plate in commercial broilers, AACs were observed to exhibit more pronounced changes in their growth plate that that in TBCs. Taken together, these results demonstrate that HIF-1α, -2α, and -3α may be important in tibial growth plate development and in the prevention of TD. The present study contributes novel insights on a therapeutic target for poultry TD.

Investigation and characterization of β-lactam resistance in Escherichia coli strains isolated from bamboo rats (Rhizomys sinensis) in Zhejiang province, China

This study was undertaken to investigate drug resistance in Escherichia coli (E. coli) strains isolated from bamboo rats in Zhejiang province of China. One hundred and fifty-four E. coli strains were isolated from dead bamboo rats. Polymerase chain reaction (PCR) was used to detect the representative genes encoding resistance to commonly used β-lactam antibiotics. Highest resistance was observed for cefradine (24.03%), followed by penicillin (20.78%) and ceftazidime (20.13%). The isolation rates of β-lactam resistance genes were 53.25, 48.70, 15.58 and 14.29% for bla TEM, bla CTX-M, bla OXA and bla SHV, respectively, while 62 (40.26%) E. coli isolates harbored multiple β-lactam resistance genes. These results also suggested that long term use of these antibiotics leads to antibimicrobial resistance. We believe that this study will provide a guideline for veterinarians and a research basis for examining resistance-encoding genes in other food animals like bamboo rats.

Molecular Detection of Indigenous Hepatitis E Virus (HEV) from Tibetan Pigs in Tibet, China

Hepatitis E is an important public health concern throughout the world. Many molecular and serological surveys have reported the prevalence and genotypic characteristics of HEV in humans and animals worldwide. However, the genotypic characterization of this virus is very limited in Tibetan pigs. Hence, we aimed to explore the genotype of HEV, prevailing among Tibetan pigs in China. For this purpose, 253 bile samples of Tibetan pigs (free-range animals) were collected from different slaughterhouses during 2017–2018 and subsequently tested for HEV RNA by RT-nPCR. A total of 11 out of 253 (4.35%) samples tested were positive for HEV RNA. Based on the sequence alignment and phylogenetic analysis, all the isolated HEV strains belonged to genotype 4 and clustered into subtype 4b by sharing more than 84.8–95.2% identities with other reported strains. Our results concluded that HEV genotype 4 is prevailing among Tibetan pigs in Tibet, China.

Effect of tetramethyl thiuram disulfide (thiram) in relation to tibial dyschondroplasia in chickens

Tetramethyl thiuram disulfide (thiram) is one of the important pesticides, which is extensively used in agriculture, but if it is combined with the cell membrane, then it causes membrane damage, bone morphogenic inactivation, and inhibited angiogenesis. Thiram has been considered a common cause of tibial dyschondrolplasia (TD) in various avian species, because it becomes the part of feed due to environmental contamination and its overuse in agriculture as pesticides or fungicide. However, there is no systematic study on the changes of the correlation indexes with toxic effect of the thiram in chickens. Therefore, we evaluated the toxic effects of thiram on growth performance of chickens, viscera organ index, pathological changes in tissue, and gene expression associated with osteoblast differentiation, vascularization, and tibial bone development. For this study, 1-day chickens (nu2009=u2009300) were randomly distributed into two equal groups, control group (normal basal diet) and thiram group (adding thiram 40xa0mg/kg in basal diet). The result presented that thiram group chickens were looking unhealthy, lazy, and showing clinical symptoms like lameness. Thiram treatment significantly reduced the performance of chickens, liver index, and tibial length compared with control group. The toxic effect of thiram increased the visceral organ index (spleen and cardiac), tibia index, and TD severity considerably. It also increased serum Ca2+ and P3+ concentration and decreased tibial density compared to control chickens but the difference was not significant. Histopathology of tibia and liver showed that there were severe lesions due to toxic effect of thiram. Furthermore, HIF-1α and VEGF antibody localizations were increased and WNT4 localization was reduced significantly in immunohistochemical analysis. This systemic study of toxic effects of thiram in chicken concluded that thiram reduced the growth performance of chickens through decreasing liver index, whereas increasing kidney, cardiac, and spleen index, and induced TD by changing the expressions of VEGF, HIF-1α, and WNT4.

Icariin: a Potential Compound for the Recovery of Tibial Dyschondroplasia Affected Chicken Via Up-Regulating BMP-2 Expression

BackgroundTibial dyschondroplasia (TD) is a skeletal disease of fast growing chicken and other avian species. It is characterized by an avascular and non-mineralized growth plate, which leads to a deformed tibial bone and lameness. Unfortunately, this disease is not only responsible for causing huge economic losses but also raises animal welfare concerns. Icariin is a flavonoid, which is isolated from Epimedium pubescens herb, and it has been used to cure different diseases including bone fractures and osteoporosis.ResultsWe designed this experiment to use icariin for the treatment of TD affect chickens; for this purpose, a total of 180 chicks were equally divided into three groups: control, TD and icariin. All the three groups were offered ad libitum same normal standard diet with an addition of thiram (50xa0mg/kg) from 3rd day to 7th day in TD and icariin group in order to induce TD in chickens. After the induction of TD, the chickens in icariin groups were fed standard diet with an addition of icariin at the rate of 10xa0mg/kg in drinking water to check the therapeutic effect of this flavonoid on TD. Our results showed that the icariin helped in restoring the TD lesion into a normal structure with significantly (Pu2009<u20090.05) up-regulating the bone morphogenetic protein-2 (BMP-2) expression in the tibial growth plates (GP).ConclusionsIcariin increased the vascular area in the growth plate and decreased the average TD score. In conclusion, this study shows that icariin is a potential compound for the recovery of TD affected chickens via up-regulating the BMP-2 expression without posing a threat of ingestion of toxic veterinary drug residues to human beings upon the consumption of treated chickens.

Physiological variations among blood parameters of domestic cats at high- and low-altitude regions of China

Abstract This study aims to determine the comparative haematological parameters and to check possible physiological changes among blood parameters of domestic cats living at high- and low-altitude regions of China. For this purpose, blood samples were randomly collected from healthy semi-long-haired Tonkinese cats in Tibet (high altitude) and Zhejiang (low altitude) regions of China. The results of present study showed that RBCs, HGB, MCH and MCHC values of Tibetan Tonkinese cats were significantly (pu2009<u2009.01) higher as compared to Zhejiang Tonkinese cats. However, MCV values of Tibetan Tonkinese cats were significantly lower (pu2009<u2009.01) as compared to Zhejiang cats. In addition, there was no significant difference among HCT values of both Tibetan and Zhejiang cats. Therefore, it was concluded that Tibetan cats had adapted the high-altitude environment of Tibet by acquiring high concentration of RBCs, HGB, MCH and MCHC, while decreased concentration of MCV content in their blood. This is the first report describing the comparative haematological parameters of Tibetan and Zhejiang Tonkinese cats living at high and low altitudes, respectively. This study provides reference values of the different blood parameters, which can be used as an important marker for the diagnosis of various diseases and utilizing in further animal research.