Dairy Cattle In China Research Articles

Study of Plasma Biochemistry and Plasma Metabolomics Differences in Montbéliard and Holstein Backcross and Holstein Heifers.

Holstein cattle are the main breed of dairy cattle in China. However, given the high degree of purebred selection of Holstein cattle, Chinese dairy cattle are increasingly being characterized by poor disease resistance, poor quality, and declining fertility. In this study, using Montbéliard × Holstein cattle as females and Montbéliard bulls as males for backcross breeding, we sought to provide a reference for improving the quality and performance of Holstein cattle and enhancing the efficiency of dairy farming. On the basis of similar physiological status and age, we selected 24 Montbéliard and Holstein backcross heifers and 11 Holstein heifers fed the same formula for comparative analyses. Plasma samples collected for plasma biochemical index analyses revealed that the content of ALB and BUN in the Montbéliard and Holstein backcross heifers was 20.83% (31.62 g/L to 26.17 g/L) and 42.36% (6.89 mmol/L to 4.84 mmol/L) higher than in the Holsteins (p < 0.01). The ALB/GLB (0.90 to 0.60, p < 0.05) was significantly higher in Montbéliard and Holstein backcross heifers than in Holstein heifers. Similarly, the activity of CAT in the backcross heifers was 61.28% (4.29 U/mL to 2.66 U/mL) higher than that in the Holstein heifers (p < 0.05). Although the activity of GSH-Px in the backcross heifers also showed an increasing trend, the difference did not reach the level of statistical significance (p = 0.052). Compared with Holstein heifers, the concentrations of IgA, IgG, and IL-4 were elevated by 32.52% (24.90 μg/mL to 18.79 μg/mL, p < 0.01), 13.46% (234.32 μg/mL to 206.53 μg/mL, p < 0.01), and 14.59% (306.27 pg/mL to 267.28 pg/mL, p < 0.05), and the contents of IL-6 and TNF-α were decreased by 15.92% (215.71 pg/mL to 256.55 pg/mL, p < 0.01) and 32.17% (7.17 ng/mL to 10.57 ng/mL, p < 0.01) in the plasma of Montbéliard and Holstein backcross heifers. Among the experimental heifers, five animals from each of the two groups were selected for plasma metabolomic analysis based on untargeted liquid chromatography-mass spectrometry. A comparison of the differential metabolites between the two heifer breeds revealed an up-regulation of d-glucuronic acid, s-glutathionyl-l-cysteine, and oleic acid levels in the backcross cattle compared with those in the Holstein heifers. We speculate that changes in the levels of these metabolites may be associated with an enhancement of the anti-inflammatory, antioxidant, and immune systems in these backcross heifers. Collectively, our findings in this study indicate that compared with 12-month-old purebred Holstein heifers, Montbéliard and Holstein backcross heifers of the same age are characterized by higher antioxidant capacity and immunity.

Study on genetic characteristics of Cryptosporidium isolates and first report of C. parvum IIdA24G2 subtype in dairy cattle in China.

Cryptosporidium is an important gastrointestinal parasite that can cause mild to severe diarrhea in various vertebrates, including humans and domestic animals. Infection is prevalent in dairy cattle, particularly calves, resulting in diarrhea and increased mortality with significant production losses. However, the prevalence and identity of Cryptosporidium spp. in cattle in Heilongjiang Province is still poorly known. Our study aimed to investigate the prevalence and species and subtype distribution of Cryptosporidium in cattle in the region. In addition, we evaluated the zoonotic potential of Cryptosporidium isolates and assessed possible transmission routes and health effects of this organism. We collected 909 fecal samples from five different farms in Heilongjiang Province between August and September 2022. The samples underwent Cryptosporidium detection by nested PCR and small subunit (SSU) rRNA gene sequence analysis. Four Cryptosporidium species were identified, including C. parvum, C. bovis, C. ryanae, and C. andersoni, with an overall prevalence of 4.4% (40/909). Based on sequence analysis of the 60 kDa glycoprotein gene of C. parvum and C. bovis, three subtypes of C. parvum were identified, namely two previously known subtypes (IIdA19G1 and IIdA20G1), and one novel subtype (IIdA24G2). Two distinct subtype families were identified in C. bovis (XXVId and XXVIe). The high diversity of Cryptosporidium in dairy cattle and the emergence of a novel subtype of C. parvum in Heilongjiang Province suggest that dairy cattle may serve as a significant source of zoonotic cryptosporidiosis infection in this region.

Tn3-like structures co-harboring of blaCTX-M-65, blaTEM-1 and blaOXA-10 in the plasmids of two Escherichia coli ST1508 strains originating from dairy cattle in China

The purpose of this study was to determine the level of horizontal transmission of the blaCTX-M-65 gene and the role of its associated mobile genetic elements (MGEs) in the bovine-derived Escherichia coli. After PCR identification, two plasmids carrying blaCTX-M-65 were successfully transferred to the recipient E. coli J53 Azr through conjugation assays and subsequently selected for Whole-Genome sequencing (WGS) analysis. The resistance profiles of these two positive strains and their transconjugants were also determined through antimicrobial susceptibility tests. Whole genome data were acquired using both the PacBio sequencing platform and the Illumina data platform. The annotated results were then submitted to the Genbank database for accession number recording. For comparison, the genetic environment of plasmids carrying the resistance gene blaCTX-M-65 was mapped using the Easyfig software. WGS analysis revealed Tn3-like composite transposons bearing blaCTX-M-65, blaTEM-1, and blaOXA-10 in the IncHI2-type plasmids of these two E. coli ST1508 strains. A phylogenetic tree was generated from all 48 assembled E. coli isolates blaCTX-M-65, blaTEM-1, and blaOXA-10 from the NCBI Pathogen Detection database with our two isolates, showing the relationships and the contribution of SNPs to the diversity between genetic samples. This study suggests that the transmissibility of blaCTX-M-65 on Tn3-like composite transposons contributes to an increased risk of its transmission in E. coli derived from dairy cattle.

Prevalence, distribution, and risk factors of bovine tuberculosis in dairy cattle in central China.

Bovine tuberculosis (bTB) is one of the priority epidemic diseases in dairy cattle in China. Continuous surveillance and evaluation of the control programs will help on improving the efficiency of bTB control policy. We designed this study to investigate both animal and herd level prevalence of bTB, as well as to determine the associated factors in dairy farms in Henan and Hubei provinces. A cross-sectional study was conducted from May 2019 to September 2020 in central China (Henan and Hubei provinces). We sampled 40 herds in Henan and six herds in Hubei via stratified systematic sampling and administrated a questionnaire consisting of 35 factors. A total of 4900 whole blood samples were collected from 46 farms, including 545 calves <six months old and 4355 cows ≥six months old. This study demonstrated a high animal-(18.65%, 95% CI: 17.6-19.8) and herd-level (93.48%, 95%CI: 82.1-98.6) prevalence of bTB in dairy farms in central China. The Least Absolute Shrinkage and Selection Operator (LASSO) and negative binomial regression models showed that herd positivity was associated with the practice of introducing new animals (RR = 1.7, 95%CI: 1.0-3.0, p=0.042), and changing the disinfectant water in the wheel bath at the farm entrance every three days or less (RR = 0.4, 95%CI: 0.2-0.8, p=0.005) which reduced the odds of herd positivity. In addition, the result illustrated that testing cows with a higher age group (≥ 60 months old) (OR=1.57, 95%CI: 1.14-2.17, p=0.006) and within the early stage of lactation (DIM=60-120 days, OR=1.85, 95%CI: 1.19-2.88, p=0.006) and the later stage of lactation (DIM≥301 days, OR=2.14, 95%CI: 1.30-3.52, p=0.003) could maximize the odds of detecting seropositive animals. Our results have plenty of benefit to improve bTB surveillance strategies in China and elsewhere in the world. The LASSO and the negative binomial regression models were recommended when dealing with high herd-level prevalence and high dimensional data in questionnaire-based risk studies.

Prevalence and multilocus genotyping of Giardia duodenalis in Holstein cattle in Yunnan, China.

Giardia duodenalis is an important zoonotic protozoon, which can infect a variety of animals, causing diarrhea and even death of animals or humans. Dairy cattle have been implicated as important sources of human G. duodenalis. However, the information about the prevalence and genetic diversity of G. duodenalis in dairy cattle in China's Yunnan Province remains limited. This study investigated the occurrence and multilocus genotyping of G. duodenalis of Holstein cattle in Yunnan Province, China. A total of 524 fresh fecal samples of Holstein cattle were randomly collected from 8 farms in Yunnan. In this study, 27.5% (144/524) of tested samples were positive for G. duodenalis infection. The highest infection ratio was found in preweaned calves (33.7%), and the infection rates of postweaned calves, growing cattle, and adult cattle were 24.5%, 23.0%, and 17.3%, respectively. The sequence analysis of SSU rRNA gene showed that the predominant assemblage of G. duodenalis in this study was assemblage E (97.9%, 141/144), whereas assemblage A was identified only in three samples (2.1%, 3/144). All G. duodenalis-positive samples were further assayed with nested polymerase chain reaction (PCR) targeting β-giardin (bg), triosephosphate isomerase (tpi), and glutamate dehydrogenase (gdh) genes, and 87, 41, and 81 sequences were obtained, respectively. Mixed infection of assemblages A and E of G. duodenalis was detected in three samples. Multilocus genotyping yielded 23 multilocus genotypes (MLGs). This is the first study that reveals the prevalence data of G. duodenalis in Holstein cattle in Yunnan Province, and the results of this study provided baseline data for the prevention and control of G. duodenalis infection in Holstein cattle in Yunnan Province, China.

Hematology Reference Intervals for Holstein Cows in Southern China: A Study of 786 Subjects.

Simple SummaryThe American Society of Veterinary Clinical Pathology guidelines recommend that hematology reference intervals (RIs) should be established based on specific animals in specific regions. However, the information on hematology RIs is still lacking for Chinese dairy cows, which account for a total number of more than 3 million. Thus, the aim of this study is to establish hematology RIs for Chinese dairy cows using a large sample size. After the collection of blood samples from 786 Holstein cows, we generated hematology RIs according to overall animals, different ages, different parities and different lactation stages. Our results provide important hematologic information for both clinicians and researchers to approach the health surveillance of Holstein cows in southern China.Hematology RIs help clinicians and researchers determine whether a hematology parameter is abnormal, which plays an important role in animal health surveillance. China is one of the largest dairy producers in the world, with millions of Holstein cows. However, there has been no published data on hematology RIs for dairy cows in China yet. Therefore, the aim of this study is to establish updated and accurate RIs for Holstein cows in southern China. To increase the accuracy of the RIs, we collected blood samples from 786 Holstein cows and analyzed 25 hematologic variables. The RIs for Holstein cows were established using the 95% percentile RIs according to the American Society of Veterinary Clinical Pathology guidelines. The effects of different ages, parities and lactation stages were also checked in this study. The data of 21, 22 and 19 out of 25 hematology parameters were significantly different between different ages, parities and lactation stages, respectively. Furthermore, the hematology RIs of separate subclasses according to different ages, parities and lactation stages were generated. Hematology RIs according to ages and lactation stages, as well as parities and lactation stages, were also assessed. Together, our results confirm that hematology RIs for cows vary by ages, parities and lactation stages. The present study helps to fill the gap in hematology RIs for Holstein cows in southern China, and our data may serve as a very useful tool for monitoring the health and welfare of dairy cattle in China.

Prevalence of bovine tuberculosis in dairy cattle in China during 2010-2019: A systematic review and meta-analysis.

BackgroundBovine tuberculosis (bTB), caused by members of the Mycobacterium tuberculosis complex bacteria, mainly Mycobacterium bovis (M. bovis), is a major threat to public health and economic development. There has been no systematic epidemiological assessment concerning bTB in dairy cattle in China.Methodology/principal findingsLiterature related to bTB in China was retrieved from China National Knowledge Infrastructure (CNKI), PubMed, ScienceDirect, VIP Chinese Journals Database, and Wan Fang Database to build the first meta-analysis for estimating the prevalence and infection moderators of bTB in dairy cattle in China. A total of 100 relevant studies published from 2010 to 2019 were included. We estimated the overall prevalence of bTB was 2.4% (95% CI: 2.1–2.8) during this decade. In the sampling year subgroup, the prevalence was lowest in 2017 or later at 0.8% (95% CI: 0.3–1.5). The lowest prevalence was 0.7% (95% CI: 0.5–1.0) in Northwestern China. The lowest prevalence was 2.1% (95% CI: 1.8–2.5) using SIT test. Heifer cows had the highest prevalence, which was 27.1% (95% CI: 9.7–49.2). The prevalence in scale farming was 3.7% (95% CI: 3.1–4.3), significantly higher than that in free-range farming (1.7%, 95% CI: 1.1–2.4). The prevalence of bTB was highest in summer at 4.0% (95% CI: 1.7–7.0). In addition, the influence of different geographical factors (altitude, longitude, latitude, precipitation, temperature, humidity) on the prevalence was analyzed.Conclusions/significanceThe results showed that bTB was widespread in China but has been gradually reduced through concerted national intervention. It is suggested that different countries should formulate corresponding prevention and control measures according to the epidemic situation in its cattle industry. Enhanced monitoring of warm and humid areas may play an important role in reducing the incidence of bTB. In addition, when large-scale breeding is promoted, attention should be paid to standardizing breeding management and improving animal welfare to reduce the prevalence of bTB in cattle.

An emerging novel bovine coronavirus with a 4-amino-acid insertion in the receptor-binding domain of the hemagglutinin-esterase gene.

The hemagglutinin-esterase (HE) protein of betacoronavirus lineage A is a secondary receptor in the infection process and is involved in the emergence of new betacoronavirus genotypes with altered host specificity and tissue tropism. We previously reported a novel recombinant bovine coronavirus (BCoV) strain that was circulating in dairy cattle in China, but this virus was not successfully isolated, and the genetic characteristics of BCoV are still largely unknown. In this study, 20 diarrheic faecal samples were collected from a farm in Liaoning province that had an outbreak of calf diarrhea (≤ 3 months of age) in November 2018, and all of the samples tested positive for BCoV by RT-PCR. In addition, a BCoV strain with a recombinant HE (designated as SWUN/A1/2018) and another BCoV strain with a recombinant HE containing an insertion (designated as SWUN/A10/2018) were successfully isolated in cell culture (TCID50: 104.25/mL and 104.73/mL, respectively). Unexpectedly, we identified the emergence of a novel BCoV variant characterized by a 12-nt bovine gene insertion in the receptor-binding domain in a natural recombinant HE gene, suggesting a novel evolutionary pattern in BCoV.Electronic supplementary materialThe online version of this article (10.1007/s00705-020-04840-y) contains supplementary material, which is available to authorized users.

Prevalence and Population Genetics Analysis of Enterocytozoon bieneusi in Dairy Cattle in China.

Enterocytozoon bieneusi, an obligate intracellular pathogen, can infect various hosts. In this study, 3527 dairy cattle fecal specimens were collected from different geographic locations in China (including 673 from Shandong province, 1,440 from Guangdong province and 1,414 from Gansu province) and examined for the presence of E. bieneusi using polymerase chain reactions targeting the ribosomal internal transcribed spacer (ITS). The dominant genotypes identified were further subtyped by multilocus sequence typing. The overall prevalence of E. bieneusi was 14.2% (501/3527), with a significant difference in prevalence among the different geographical locations (P < 0.001). Our logistic regression analysis showed that all four variables (farming model, location, age, and clinical manifestations) had strong effects on the risk of contracting E. bieneusi. Sequence analysis revealed 11 genotypes: eight known genotypes (J, I, BEB4, BEB10, D, EbpC, CM19, and CM21) and three novel genotypes (named here as CGC1, CGC2, and CGC3). Genotypes J and I, the commonest, were found on all farms across the three provinces. Our linkage disequilibrium analysis showed a clonal population structure in the E. bieneusi dairy cattle population but the ITS genotypes had different population structures. Phylogenetic and haplotype network analysis showed the absence of geographical segregation in the E. bieneusi dairy cattle populations. Instead, they revealed the presence of host adaptation to the E. bieneusi populations in various animals. Our findings augment the current understanding of E. bieneusi transmission dynamics.

Prevalence of a novel bovine coronavirus strain with a recombinant hemagglutinin/esterase gene in dairy calves in China.

Bovine coronavirus (BCoV) is the causative agent of diarrhoea in newborn calves, winter dysentery in adult cattle and respiratory tract illnesses in cattle across the world. In this study, a total of 190 faecal samples from dairy calves with diarrhoea were collected from 14 farms in six Chinese provinces, and BCoV was detected in 18.95% (36/190) of the samples by reverse transcriptase polymerase chain reaction. Full‐length spike, hemagglutinin/esterase (HE), nucleocapsid and transmembrane genes were simultaneously cloned from 13 clinical samples (eight farms in four provinces), and most of the BCoV strains showed a unique evolutionary pattern based on the phylogenetic analysis of these genes. Interesting, 10 of the 13 strains were identified as HE recombinant strains, and these strains had experienced the same recombination event and carried the same recombination sites located between the esterase and lectin domain. They also shared an identical aa variant (F181V) in the R2‐loop. Moreover, 9/10 strains displayed another identical aa variant (P, S158A) in the adjacent R1‐loop of the HE gene, which differs from the other available BCoV HE sequences in the GenBank database. Our results showed that BCoV is widely circulating in dairy cattle in China, contributing to the diagnosis and control of dairy calves diarrhoea. Furthermore, a BCoV strain that carries a recombinant HE gene has spread in dairy calves in China. To the best of our knowledge, this is the first description of an HE recombination event occurring in BCoV; this is also the first description of the molecular prevalence of BCoV in China. Our findings will enhance current understanding about the genetic evolution of BCoV.

Prevalence of Cryptosporidium in dairy cattle in China during 2008–2018: A systematic review and meta-analysis

Cryptosporidium is the causative agent of cryptosporidiosis. Cryptosporidium not only has a worldwide distribution, but also can infect various hosts, including dairy cattle and humans. Although numerous researches on Cryptosporidium infection in cattle have been conducted, no nationwide study on the prevalence of Cryptosporidium infection in dairy cattle in mainland China was carried out. In this meta-analysis, five databases, including PubMed, ScienceDirect, China National Knowledge Infrastructure (CNKI), Chongqing VIP, and Wanfang, were used to search for published papers regarding Cryptosporidium infection in dairy cattle in China from inception to February 25, 2019. Our study obtained 60 eligibility studies that reported Cryptosporidium infection in dairy cattle. We estimated the pooled Cryptosporidium prevalence to be 17.0% (3,901/33,313), with 16.9% (722/5,191) in Central China, 17.4% (959/6,162) in Eastern China, 29.8% (404/2,021) in Northeastern China, 15.7% (227/2,344) in Northern China, 15.8% (1,042/11,452) in Northwestern China, 9.5% (494/5,758) in Southern China, and 13.7% (53/385) in Southwestern China. The pooled prevalence of Cryptosporidium in before 2000 group (28.0%, 944/3,417) was significantly higher than in 2000–2010 group (11.1%, 384/3,643) and after 2010 group (13.7%, 2,134/22,411). Cattle with the age of ≤ 12 months (22.5%, 2,142/12,948) had a significantly higher prevalence than those of > 12 months (9.5%, 840/10,282). The pooled prevalence of Cryptosporidium in different seasons ranged from 8.2% (343/4,552) in Autumn to 19.5% (285/1,570) in Winter. Diarrhea cattle (38%, 133/477) had a higher Cryptosporidium prevalence than non-diarrhea cattle (13.0%, 367/2423). The pooled prevalence of Cryptosporidium in different provinces was various, with the highest (35.6%, 355/1,667) in Heilongjiang province, and the lowest (4.3%, 15/440) in Tianjin province. The univariate meta-regression analyses indicated that the collection year (P = 0.002) and age of cattle (P = 0.001) might be sources of heterogeneity. This systematic review suggests that China is a country where cryptosporidiosis frequently occurs in cattle. Due to the particular relationship between dairy cattle and feeder, further research is required to investigate the links between cattle ownership and Cryptosporidium infection.

Short communication: Genotyping and single nucleotide polymorphism analysis of bovine leukemia virus in Chinese dairy cattle

Bovine leukemia virus (BLV) causes enzootic leucosis in cattle and is classified into 10 genotypes with a worldwide distribution, except for several European countries, Australia, and New Zealand. Although BLV is widespread in Chinese cows with the positive rate of 49.1% at the individual level, very little is known about the BLV genotype in dairy cattle in China. To determine BLV genetic variability in cows in China, 112 BLV-positive samples from 5 cities in China were used for BLV molecular characterization in this study. Phylogenetic analysis using the neighbor-joining method on partial env sequence encoding gp51 obtained from 5 Chinese cities and those available in GenBank (n = 53, representing BLV genotype 1-10) revealed the Chinese strains belonged to genotype 6. Seven unique SNP were identified among Yancheng, Shanghai, and Bengbu strains out of the total 12 SNP identified in Chinese strains. The genotyping coupled with SNP analysis of BLV can serve as a useful molecular epidemiological tool for tracing the source of pathogens. This study highlights the importance of genetic analysis of geographically diverse BLV strains to understand BLV global genetic diversity.

Brucellosis seroprevalence in dairy cattle in China during 2008–2018: A systematic review and meta-analysis

Brucellosis remains one of the most common zoonotic diseases globally with more than a half million human cases reported annually. The Brucella reservoir associated with livestock brucellosis poses a significant threat to public health, and awareness of the seroprevalence and spatial distribution of livestock brucellosis is valuable for the prevention and control of diseases caused by Brucella, especially human brucellosis. Therefore, we conducted a systematic review and meta-analysis to evaluate the seroprevalence of brucellosis in dairy cattle in China. We retrieved 88 studies related to the seroprevalence of brucellosis in dairy cattle in China in which samples were harvested between 2008 and 2018. The results of our systematic review and meta-analysis reveal that the overall seroprevalence of brucellosis in dairy cattle herds in China was 1.9% during the selected period, rising from 1.6% in 2008–2012 to 2.6% in 2013–2018. In Northern China, where the traditional agropastoral areas with more developed animal breeding industry are located, the brucellosis seroprevalence was >10%. In contrast, the seroprevalence of brucellosis in Southern China reached only 5.5%. At the provincial level, the highest brucellosis seroprevalence in dairy cattle was estimated at >30% in Jilin province, followed by Shanxi, Ningxia, Inner Mongolia, and Guizhou, each with a prevalence of 10–20%. Additionally, the seroprevalence of brucellosis in some local areas was >30% or even >50%, indicating that Brucella infection was highly endemic in dairy herds in China. Our data may facilitate the prevention and control of brucellosis in domestic animals in China. Further epidemiological surveillance and the administration of a comprehensive monitoring program to determine the risk factors associated with brucellosis incidence in humans and domestic animals are recommended to refine brucellosis control strategies.

A systematic review and meta-analysis of the epidemiology of bovine viral diarrhea virus (BVDV) infection in dairy cattle in China

Bovine viral diarrhea virus (BVDV) infection causes significantly economic losses to cattle industry worldwide, also including China. The epidemiological prevalence of infection associated with BVDV in dairy cattle has not been systematically assessed in China. Therefore, we undertook this study to evaluate prevalent of BVDV infection. We conducted a systematic review and meta-analysis of data from papers on the BVDV incidence and prevalence in dairy cattle in China by searching China Science and Technology Journal Database, China National Knowledge Infrastructure (CNKI), Wan Fang Database and PubMed for publication from March 2003 to March 2018. The 41 studies reporting the prevalence of BVDV in cattle in China were selected upon our inclusion criterion. The pooled BVDV prevalence in dairy cattle in China was estimated to 53.0% (95% CI 40.2–65.7) based on the data obtained from the 27,530 cows tested using serological or virological assay in the qualified papers published during the periods (χ² = 51,861.0, I2 = 99.9%). The highest BVDV positive rate in dairy flocks reached 90.0% in Fujian province of China, followed by Shaanxi (88.9%) and Shandong (83.3%). The prevalence in the six administrative districts of China was validated to be highly variable (25.7%–72.2%) and reached 72.2% in dairy cattle flocks of Northern China. Besides, the BVDV-RNA positive rate was estimated 27.1% (95% CI 17.3–37.0) based on 6 studies, comparatively, the pooled BVDV seroprevalence based on 35 studies was about 57.0% (95% CI 44.4–69.5) in China. This systematic review and meta-analysis firstly established an estimated prevalence of BVDV in dairy herds in China, indicating that the BVDV infection is escalating, though there is a bias in the number of studies between 2003–2009 and 2010–2018 timescales. This study may help understand the status of BVDV infection in dairy herds in China. Further extensive and comprehensive investigation is recommended, and effective intervention measures for preventing and controlling BVDV spread in dairy herds should be deployed, especially herds that have been exposed to BVDV.

The isolation and molecular characterization of Mycobacterium avium subsp. paratuberculosis in Shandong province, China.

BackgroundMycobacterium avium subspecies paratuberculosis (Map) causes Johne’s disease in domestic and wild ruminants. It has been a debate that whether Map can cause Crohn’s disease in human. To our knowledge there is no report about molecular characterization of Map in China, although several Map strains have been reported in other country. The objectives of this study was to know the recent prevalence of Johne’s disease in dairy farms in Shandong province, and have a better understanding of genotypic distribution of Map in China.MethodsJohne’s disease was detected from 1038 individuals in 19 dairy farms by ELISA. Map in fecal and milk specimens was identified by Ziehl-Neelsen staining and confirmed using PCR-REA. In addition, frozen sections of ileum and mesenteric lymph nodes from two Map shedding cows were performed to observe the histopathological changes. Next-generation sequencing technology was performed to get whole genome sequences.ResultA total of 121 (11.7 %) animals were positive for Map antibody from 1038 sera tested, and 11 (57.9 %) dairy herds were positive for Map antibody. Typically histopathologic changes were observed in mesenteric lymph nodes. We have successfully isolated two Map strains, which both were Map-C. The current genome-wide analysis showed that the genome size of our isolates are respectively 4,750,273 and 4,727,050 bp with a same G + C content of 69.3 %, and the numbers of single nucleotide polymorphisms (SNPs) against Map K-10 are respectively 292 and 296.ConclusionMap is a prevalent pathogen among dairy cattle in China. This study successfully isolated two Map strains from one Chinese dairy herd with signs of diarrhoea, and identified that the two isolates were both Map-C. Furthermore, these isolates were most closely related to Map K-10.

High seroprevalence of Coxiella burnetii in dairy cattle in China

Coxiella burnetii is the agent of Q fever, a zoonosis which occurs worldwide. As there is little reliable data on the organism in China, we investigated C. burnetii infections in dairy cattle herds around the country. Opportunistic whole blood samples were collected from 1140 dairy cattle in 19 herds, and antibodies to phase I and II C. burnetii antigens were detected using commercial ELISA kits. Seropositive cattle (381/1140, 33%) were detected in 13 of the 15 surveyed provinces and in 16 of the 19 herds (84%) studied. Our data indicates C. burnetii is widespread in China and that animal and human health workers should be aware of the possibility of Q fever infection in their patients.

Seroprevalence and correlates of Toxoplasma gondii infection in dairy cattle in northwest China.

Prevalence of antibodies to Toxoplasma gondii and risk factors with infection were assessed in dairy cattle from Gansu Province and Ningxia Hui Autonomous Region (NXHAR), northwest China. In total, 1657 serum samples were collected and assayed by the modified agglutination test. The overall seroprevalence was 4.83% at a 1:100 cut-off, with titers of 1:100 in 72, 1:200 in 4, 1:400 in 4. Among the risk factors examined, no statistically significant difference was observed between T. gondii seroprevalence and regions or age of dairy cattle in the logistic regression analysis (P>0.05) and left out of the final model. However, numbers of pregnancies of dairy cattle was considered as main risk factor associated with T. gondii infection. Dairy cattle in nulliparity group (8.89%) had 6 times (OR=6.31, 95% CI, 2.69-14.83, P<0.001) higher risk of being seropositive compared to dairy cattle in 3 or above 3 pregnancies group (1.52%), followed by 1 pregnancy group (4.27%) had nearly 3 times (OR=2.89, 95% CI, 1.11-7.52, P = 0.03) higher risk of being seropositive compared to dairy cattle in 3 or above 3 pregnancies group, although no statistical difference was found between 2 pregnancies group and 3 or above 3 pregnancies group (P = 0.70). The results of this survey indicated the presence of T. gondii infection in dairy cattle in Gansu Province and NXHAR, which enriches the epidemiological data of T. gondii infection in dairy cattle in China, and is helpful to strengthen prevention and control of T. gondii infection in dairy cattle in these two regions.

A bovine viral diarrhea virus type 1a strain in China: isolation, identification, and experimental infection in calves

BackgroundBovine viral diarrhea virus (BVDV) is one of the most important pathogens in cattle. Previously, BVDV sub-genotypes of 1b, 1c, 1d, and 1 m were detected in China. However, isolation of BVDV type 1a from cattle has not been reported in China. In 2010, twenty nasal swabs and blood samples were collected from the cattle suspected BVDV infection in Henan province, China. A BVDV isolate was isolated using cell culture, and the pathogenesis of the virus isolate was studied.MethodsVirus isolation was performed on MDBK cells. The virus identification was conducted by RT-PCR, neutralization test and immunofluorescence assay. In order to determine the genotype of the newly isolated virus, the 5′ un-translated region (5′UTR) of the virus isolate was cloned, sequenced and phylogenetically analyzed. To evaluate the virulence of the virus isolate, four BVDV sero-negative calves were intranasally inoculated with the virus suspension. Rectal temperatures and clinical signs were recorded daily. Blood samples were analyzed for changes in white blood cell counts, and tissue samples were taken for histopathology analysis.ResultsA new isolate of bovine viral diarrhea virus (BVDV), named HN01, was isolated from the nasal swabs using MDBK cell culture. The HN01 strain caused cytopathic effect (CPE) in MDBK cell cultures after two passages. The virus specifically reacted to BVDV1-specific monoclonal antibody in an immunofluorescence assay. A fragment of 288 bp of genome from this isolate was amplified by the RT-PCR. Phylogenetic analysis of 5′UTR indicated that the virus was BVDV 1a. In the pathogenesis study, four calves experimentally infected with the BVDV strain developed depression, cough and other clinical signs. Calves showed high temperature over 40°C, and white blood cell counts dropped more than 40%.ConclusionsA new subgenotype 1a strain of BVDV was firstly isolated from dairy cattle in China. The experimental infection showed that the virus was moderate pathogenic to cattle and can be used as a BVDV challenge virus to evaluate the efficacy of BVDV vaccines in the target animals.

Cryptosporidium andersoni is the predominant species in post-weaned and adult dairy cattle in China

Dairy industry plays an important role in the agricultural economy of China. To estimate the prevalence and public health significance of cryptosporidiosis in post-weaned and adult dairy cattle in China, during four consecutive years (from 2006 to 2009), a total of 1315 fecal samples from 22 dairy cattle farms in ten prefectures in Henan Province were examined for the presence of Cryptosporidium oocysts. The overall prevalence of Cryptosporidium was 7.9%, with the highest infection rate (11.3%) in 3 to 11-month-old calves and the lowest infection rate (1.0%) in >2-year-old cows (p<0.01). Cryptosporidium-positive samples (n=104) were analyzed by PCR-restriction fragment length polymorphism (RFLP) analysis of the small subunit (SSU) rRNA gene, and 25 representative samples were further analyzed by DNA sequencing of the PCR products. Cryptosporidium bovis and Cryptosporidium andersoni were identified. C. andersoni (84/104) was the predominant species and was found in all age groups, whereas C. bovis (20/104) was only detected in 3 to 11-month-old calves. Thus, C. andersoni appears to be the dominant species in weaned dairy calves and heifers in China, in contrast with its common occurrence in adult cattle in other parts of the world.