Houqiang Luo

Tibial dyschondroplasia is highly associated with suppression of tibial angiogenesis through regulating the HIF-1α/VEGF/VEGFR signaling pathway in chickens

Tibial dyschondroplasia (TD) is an intractable poultry problem that is characterized by the appearance of non-vascularized and non-mineralized cartilage masses in tibial growth plates (TGPs). However, the role of angiogenesis inhibition in the occurrence of TD remains unknown. In this study, we found that, compared to low-altitude Arbor Acres chickens (AACs), high-altitude Tibetan chickens showed higher tibial vascular distributions that were accompanied by up-regulation of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor A (VEGFA) and VEGF receptors. These observations provide insights into hypoxia-induced angiogenesis, which may be related to the absence of TD in high-altitude native Tibetan chickens. Importantly, hypoxia experiments also showed that during hypoxia, tibial angiogenesis was enhanced, which was due to pro-angiogenic factor up-regulation (including VEGFA, VEGFR1, VEGFR2, and IL-8), in AACs. Moreover, we observed that thiram-induced TD could strongly inhibit tibial angiogenesis in the hypertrophic zone through coordinated down-regulation of HIF-1α and pro-angiogenic factors, leading to a disruption in the blood supply to the TGP. Taken together, these findings reveal that the occurrence of TD is highly associated with inhibition of tibial angiogenesis through down-regulated expression of HIF-1α, VEGFA and VEGF receptors, which results in suppression of TGP development.

First report of Metastrongylus pudendotectus by the genetic characterization of mitochondria genome of cox1 in pigs from Tibet, China

Lungworms, a world wild distributed parasites cause serious respiratory diseases to the pigs. A high infection rate of Metastrongylus lungworms has been found in Tibetan pigs being slaughtered in different slaughter houses of Tibet autonomous region. The main aim of our study was to detect and confirm the lungworm parasite by the genetic characterization of mitochondrial cox1genome isolated from the lungs of Tibetan pig. The adult lungworms were collected from the lungs of slaughtered pigs and identification was done through morphological examinations. Total genomic DNA of the extracted worms was performed and a fragment (∼450bp) of the cox1 of mitochondrial (mt) gene was amplified. Amplicons were cloned into PGEM(®)-T Easy vector and the positive clones were sequenced from a commercial company. Sequence and phylogenetic analysis were performed by software of DNAMAN and MEGA respectively. The results revealed that the lungworms infecting the Tibetan pigs were Metastrongylus pudendotectus (M. pudendotectus). To our knowledge, this is the first report for the isolation and identification for the genetic characterization of mitochondria (mt) genome of cox1 of M. pudendotectus derived from Tibetan pigs in Tibet, China.

Effect of tetramethylpyrazine on tibial dyschondroplasia incidence, tibial angiogenesis, performance and characteristics via HIF-1α/VEGF signaling pathway in chickens.

Tibial dyschodroplasia (TD) is a most common pathological condition in many avian species that is characterized by failure of growth plate (GP) modeling that leads to the persistence of avascular lesion in the GP. Tetramethylpyrazine (TMP) is widely used to treat neurovascular disorders and pulmonary hypertension, but no report is available about promoting effect of TMP against TD. Therefore, a total of 210 broiler chicks were equally divided into three groups; Control, TD and TMP. During the experiment mortality rate, chicken performance indicators (daily weight, average daily feed intake, average daily weight gain and feed conversion ratio), tibia bone indicators (weight, length, width of tibial and the size of GP) in addition to gene expression of HIF-1α and VEGF were examined. The results showed that TMP administration restore the GP width, increase growth performance, and mitigated the lameness in broiler chickens. The expression of HIF-1α and VEGF increased significantly in TD affected thiram induced chicks. Whereas, TMP treatment down-regulated HIF-1α and VEGF genes and proteins expressions. The present study demonstrates that the TMP plays an important role in angiogenesis during the impairment and recovery of GP in TD via regulation of the HIF-1α/VEGF signaling pathway in chickens.

Prevalence, Associated Risk Factors, and Phylogenetic Analysis of Toxocara vitulorum Infection in Yaks on the Qinghai Tibetan Plateau, China.

Toxocara vitulorum has been rarely reported in yaks at high altitudes and remote areas of Sichuan Province of Tibetan Plateau of China. The current study was designed to investigate the prevalence, associated risk factors, and phylogenetic characteristics of T. vitulorum in yak calves on the Qinghai Tibetan plateau. Fecal samples were collected from 891 yak calves and were examined for the presence of T. vitulorum eggs by the McMaster technique. A multivariable logistic regression model was employed to explore variables potentially associated with exposure to T. vitulorum infection. T. vitulorum specimens were collected from the feces of yaks in Hongyuan of Sichuan Province, China. DNA was extracted from ascaris. After PCR amplification, the sequencing of ND1 gene was carried out and phylogenetic analyses was performed by MEGA 6.0 software. The results showed that 64 (20.1%; 95% CI 15.8–24.9%), 75 (17.2; 13.8–21.1), 29 (40.9; 29.3–53.2), and 5 (7.6; 2.5–16.8) yak calves were detected out to excrete T. vitulorum eggs in yak calve feces in Qinghai, Tibet, Sichuan, and Gansu, respectively. The present study revealed that high infection and mortality by T. vitulorum is wildly spread on the Qinghai Tibetan plateau, China by fecal examination. Geographical origin, ages, and fecal consistencies are the risk factors associated with T. vitulorum prevalence by logistic regression analysis. Molecular detection and phylogenetic analysis of ND1 gene of T. vitulorum indicated that T. vitulorum in the yak calves on the Qinghai Tibetan plateau are homologous to preveiously studies reported.

Epidemiologic Survey of Japanese Encephalitis Virus Infection, Tibet, China, 2015.

We investigated Japanese encephalitis virus (JEV) prevalence in high-altitude regions of Tibet, China, by using standard assays to test mosquitoes, pigs, and humans. Results confirmed that JEV has spread to these areas. Disease prevention and control strategies should be used along with surveillance to limit spread of JEV in high-altitude regions of Tibet.

Assessment of Serum Trace Elements in Diarrheic Yaks ( Bos grunniens ) in Hongyuan, China

Diarrhea is one of the main causes of animal deaths especially in neonatal calves. The objective of this study was to investigate the blood serum for cobalt (Co), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), Molybdenum (Mo), selenium (Se), and zinc (Zn) concentrations in yak serum samples by inductively coupled plasma mass spectrometry (ICP-MS) from 35 yaks consisting of 10 healthy and 25 diarrheic ones. Our observations indicated that the concentrations of Mn, Zn, and Fe in yak affected with diarrhea had statistically significant reduced levels as compared to the control group (P < 0.05). The mean concentration of Cu in blood serum of diarrheic yaks was found significantly higher than the normal ones (P < 0.05). The Mn and Zn in the serum of diarrheic groups were observed significantly different (P < 0.05) with different ages. These findings can provide for some valuable information for the investigation on relationship between trace elements in the serum of normal and diarrheic yaks.

Prevalence of three Oesophagostomum spp. from Tibetan Pigs analyzed by Genetic Markers of nad1, cox3 and ITS1

Parasitic nematodes of Oesophagostomum spp., commonly known as ‘nodular worms’ are one of the most widely distributed and prevalent emerging zoonotic nematodes. However, little is known about the prevalence and gene characteristics of those parasites in Tibetan pigs. Therefore, a study was carried out to investigate the prevalence, isolation and identification of Oesophagostomum spp from Tibetan pigs by genetic markers of nad1, cox3 and ITS1 for the first time. The results revealed that the infection rate of O. dentatum and O. quadrispinulatum by genetic markers of nad1 was 81.13%; 35 (66.04%); the O. dentatum and O. quadrispinulatum by genetic markers cox3 was 66.04%, and O. dentatum and O. stephanostomum by genetic markers ITS1 was found to be 77.36%. Interestingly, the O. stephanostomum specie was identified and isolated from 90.48% stomach and 69.23% colon samples by genetic markers of ITS1. The present study, for the first time has described the presence and genetic characterization of Oesophagostomun spp of O. dentatum, O. quadrispinulatum and especially O. stephanostomum in Tibetan pigs from the high and remote Tibetan plateau. A public concern should be raised in terms of economical losses and severe public health problem.

Seroprevalence of Toxoplasma gondii infection in zoo and domestic animals in Jiangxi Province, China

Toxoplasma gondii is a zoonotic protozoan parasite that infects a wide range of warm-blooded animals throughout the world. In the present study, antibodies to T. gondii were determined using a commercial indirect hemagglutination (IHA) test in wild animals in a zoo. Three of 11 giraffes (Giraffa camelopardalis) (27%), 1 of 5 wolves (Canis lupus laniger) (20%), 1 of 6 hippopotamuses (Hippopotamus amphibious) (17%), and 2 of 9 tundra swans (Cygnus columbianus) (22%) were found to be positive. No antibodies were detected in leopards (Panthera pardus), wild geese (Anser cygnoides), and Eastern grey kangaroos (Macropus giganteus). Domestic species from 13 counties of Jiangxi Province, China were also investigated by an indirect hemagglutination (IHA) test. Thirty-five of 340 goats (10%), 94 of 560 water buffaloes (17%), and 4 of 35 cattle (11%) were found to be seropositive. This is the first report of T. gondii infection in animals kept in zoos and domestic animals in this province.

Epidemiology of Toxoplasma gondii infection in native Tibetans in Tibet, China.

Toxoplasma gondii (T. gondii) is one of the most common and widespread protozoan parasite which caused toxoplasma infection in animals and humans. Toxoplasma infection in healthy people is usually asymptomatic and it can lead to serious pathological effects in congenital cases and immunodeficient patients. The aim of the study is to investigate the prevalence of T. gondii infection in native Tibetans in Tibet as scarce information is available on such a high plateau. A total of 600 blood samples were collected from Tibetans people who visited the hospital in Nyingchi city with the history of flu, swollen lymph glands, muscle pain, and neurological or eyes complications (showing single or more signs). The serum of collected samples were tested for IgG antibody by ELISA. The result showed that the prevalence in men and women were 11.3% and 11.0%, respectively, the prevalence in different ages were 9.8% in <7 years, 6.2% in ≥7–17 years, 10.1% in ≥18–40 years, 14.8% in ≥41–65 years and 11.6% in ≥66 years. The current study indicates the first report of this zoonotic parasite infection in Tibetan people.

Analysis of the internal transcribed spacer region of Ascaris suum and Ascaris lumbricoides derived from free range Tibetan pigs.

Abstract Ascaris suum (A. suum) is the most commonly occurring worldwide internal parasite of pigs; however, little is known about this organism in Tibetan pigs in China. A study was carried to isolate and identify the characteristics of internal transcribed spacer region (ITS) gene of A. suum derived from Tibetan pigs. Adult nematodes were collected from Tibetan pigs in 2015–2016. Total genomic DNA of the extracted parasites was performed and a fragment of the ITS of mitochondrial (mt) gene was amplified. The amplicons were cloned into PGEM®-T Easy Vector (Promega, WI) and the positive clones were sequenced by ABI 3730 × 1 sequencer. The sequence and phylogenetic analysis were performed by ClustalWVer. 1.4 and MEGA 6.0 software, respectively. Results indicated that the identity of A. suum isolates was 98.4%–99.9% with previously reported pig isolates, and 99.4%–99.7% with A. lumbricoides isolates. To the best of our knowledge, this is the first report describing the characteristics of ITS gene of A. suum derived from the Tibetan pigs from high and remote areas depicting high identity with the isolates of both A. suum and A. lumbricoides.