Molecular epidemiology, characterization of virulence factors and antibiotic resistance profile of Streptococcus agalactiae isolated from dairy farms in China and Pakistan

Streptococcus agalactiae is one of the most common pathogens that cause bovine mastitis worldwide. Identifying pathogen prevalence and virulence factors is critical for developing prevention and control approaches. Herein, 1 161 milk samples from various dairy farms in China(n=558) and Pakistan(n=603) were collected between 2019–2021 and were subjected to S. agalactiae isolation. Prevalence, serotyping, virulence genes, and antibiotic-resistant genes of S. agalactiae were evaluated by PCR assay. All isolates were characterized for haemolysis, biofilm production, cytotoxicity, adhesion, and invasion on bovine mammary epithelial cells. The prevalence of S. agalactiae-induced mastitis in cattle was found to be considerably higher in Pakistan than in China. Jiangsu and Sindh provinces had the highest area-wise prevalence in China and Pakistan, respectively. Serotypes Ia and II were prevalent in both countries, whereas serotype III was found only in Pakistan. Moreover, all isolates tested positive for PI-2b gene but negative for PI-1 and PI-2a genes. All isolates harboured cfb, cylE, hylB, and fbsB virulent genes, whereas many of them lacked bibA, rib and bca. However, the absence of bac and scp genes in Chinese isolates and cspA in Pakistani isolates was noted, while spb1 and lmb were not detected in isolates of both countries. Pakistani isolates, particularly serotype Iapositive, had a considerably higher ability to produce biofilm, haemolysis, cytotoxicity, adhesion, and invasion than Chinese isolates. Most of the isolates were phenotypically resistant to tetracycline, erythromycin, and clindamycin and genotypic resistance was confirmed by the presence of ermA, ermB, tetM and tetO genes. Our study highlights the antimicrobial resistance profile and virulence-related factors contributing to the epidemiological spread of mastitis-causing S. agalactiae in China and Pakistan. The findings may facilitate future studies designed to develop improved treatment and control strategies against this pathogen.

Molecular pathological epidemiology in diabetes mellitus and risk of hepatocellular carcinoma

Molecular pathological epidemiology(MPE) is a multidisciplinary and transdisciplinary study field,which has emerged as an integrated approach of molecular patho-logy and epidemiology,and investigates the relationship between exogenous and endogenous exposure factors,tumor molecular signatures,and tumor initiation,progression,and response to treatment.Molecular epidemiology broadly encompasses MPE and conventional-type molecular epidemiology.Hepatocellular carcinoma(HCC) is the third most common cause of cancer-associated death worldwide and remains as a major public health challenge.Over the past few decades,a number of epidemiological studies have demonstrated that diabetes mellitus(DM) is an established independent risk factor for HCC.However,how DM affects the occurrence and development of HCC remains as yet unclearly understood.MPE may be a promising approach to investigate the molecular mechanisms of carcinogenesis of DM in HCC,and provide some useful insights for this pathological process,although a few challenges must be overcome.This review highlights the recent advances in this field,including:(1) introduction of MPE;(2) HCC,risk factors,and DM as an established independent risk factor for HCC;(3) molecular pathology,molecular epidemiology,and MPE in DM and HCC;and(4) MPE studies in DM and risk of HCC.More MPE studies are expected to be performed in future and I believe that this field can provide some very important insights on the molecular mechanisms,diagnosis,personalized prevention and treatment for DM and risk of HCC.

Molecular epidemiology and phylogeny of Nipah virus infection:A mini review

Nipah virus(Ni V) is a member of the genus Henipavirus of the family Paramyxoviridae,characterized by high pathogenicity and endemic in South Asia.It is classified as a Biosafety Level-4(BSL-4) agent.The case-fatality varies from 40%-70% depending on the severity of the disease and on the availability of adequate healthcare facilities.At present no antiviral drugs are available for Ni V disease and the treatment is just supportive.Phylogenetic and evolutionary analyses can be used to help in understanding the epidemiology and the temporal origin of this virus.This review provides an overview of evolutionary studies performed on Nipah viruses circulating in different countries.Thirty phylogenetic studies have been published from 2000 to 2015 years,searching on pub-med using the key words ‘Nipah virus AND phylogeny' and twenty-eight molecular epidemiological studies from 2006 to 2015 have been performed,typing the key words ‘Nipah virus AND molecular epidemiology'.Overall data from the published study demonstrated as phylogenetic and evolutionary analysis represent promising tools to evidence NiV epidemics,to study their origin and evolution and finally to act with effective preventive measure.

Molecular Epidemiology of Enteroviruses Associated with Hand, Foot, and Mouth Disease in the Mainland of China

Hand, foot, and mouth disease （HFMD） is a common contagious illness which occurs worldwide both sporadically and in epidemics. The disease mainly affects, children and the typical symptoms, which may resolve spontaneously, include mucocutaneous papulovesicular lesions on the hands, feet, mouth, and buttocks. In rare cases, however, the patients may also develop neurological complications such as neurogenic pulmonary edema,

Characterization of Avian Influenza A(H7N9) Virus Prevalence in Humans and Poultry in Huai′an,China：Molecular Epidemiology,Phylogenetic,and Dynamics Analyses

Objective To trace the source of human H7N9 cases in Huai＇an and elucidate the genetic characterization of Huai＇an strains associated with both humans and birds in live poultry market.Methods An enhanced surveillance was implemented when the first human H7N9 case was confirmed in Huai＇an.Clinical specimens,cloacal swabs,and fecal samples were collected and screened by real-time reverse transcription-polymerase chain reaction（RT-PCR） for H7N9 virus.The positive samples were subjected to further RT-PCR and genome sequencing.The phylodynamic patterns of H7N9 virus within and separated from Huai＇an and evolutionary dynamics of the virus were analyzed.Results Six patients with H7N9 infection were previously exposed to live poultry market and presented symptoms such as fever（〉38.0 °C） and headaches.Results of this study support the hypothesis that live poultry markets were the source of human H7N9 exposure.Phylogenetic analysis revealed that all novel H7N9 viruses,including Huai＇an strains,could be classified into two distinct clades,A and B.Additionally,the diversified H7N9 virus circulated in live poultry markets in Huai＇an.Interestingly,the common ancestors of the Huai＇an H7N9 virus existed in January 2012.The mean nucleotide substitution rates for each gene segment of the H7N9 virus were（3.09-7.26）×10-3 substitutions/site per year（95% HPD：1.72×10-3 to 1.16×10-2）.Conclusion Overall,the source of exposure of human H7N9 cases in Huai＇an was live poultry market,and our study highlights the presence of divergent genetic lineage of H7N9 virus in both humans and poultry specimens in Huai＇an.

Molecular Epidemiology and Sequencing of the G-L Intergenic Region of Rabies Viruses Isolated in China

一组 25 个狂犬病病毒(RABV ) ，从 24 条狗和一个人的盒子恢复了，在在 2004 和 2006 之间的中国从各种各样的区域被收集。G-L intergenic 区域的基因、种系发生的分析在 25 街 RABV 被执行孤立， CTN 疫苗 7 拉紧代。学习基于 519 bp 核苷酸顺序的比较，包含 G-L intergenic 区域。中国街紧张的核苷酸顺序相同从 95.5% ～ 100% 。种系发生的分析证明中国的所有孤立清楚地在 Lyssavirus 遗传型 1 支持了所有中国病毒的放置，他们根据他们的地理起源是分布式的。所有仔细中国紧张被联系，但是他们能仍然被划分成二个组：一些街紧张和一些 CTN 紧张。这研究基于 G-L Intergenic 区域的序列关于狂犬病病毒的分子的传染病学介绍细节。关键词狂犬病病毒 - 分子的传染病学 - G-L intergenic 区域 - 中国 CLC 数字 R373.33 基础条款：第 10 国家 five-year-plan (2004BA718 b03 ) 的关键技术 R&D

Molecular Epidemiology of Coxsackievirus B1-5 Associated with HFMD in Fujian Province, China, 2011-2016

Hand,foot and mouth disease(HFMD)is a common infectious disease that usually affects children less than 5 years of age.HFMD is caused by human enteroviruses(HEVs).HEVs,members of the Enterovirus genus of the Picornaviridae(small RNA virus)family.

Neosporosis: An Overview of Its Molecular Epidemiology and Pathogenesis

Neospora caninum (N. caninum), a cyst-forming protozoan parasite, is a major cause of bovine abortions and neonatal mortality worldwide. N. caninum has a broad intermediate host range, and its sexual cycle occurs exclusively in canids. Another species of Neospora, Neospora hughesi (N. hughesi), has been identified and causes myeloencephalitis in horses. Although molecular epidemiology studies are in their infancy, the 18S ribosomal RNA (rRNA) and ITS1 regions within the small subunit ribosomal RNA (ssuRNA) and an N. caninum species-specific DNA probe (pNc5) have been used extensively to differentiate Neospora from other closely related apicomplexan parasites. While these repetitive regions have higher sensitivity and specificity than housekeeping or antigen genes, they suffer from low discriminatory power and fail to capture intra-species diversity. Similarly, although multiple minisatellite or microsatellite marker studies have shown clear geographic substructures within Neospora, strains are often misclassified due to a convergence in the size of different alleles at microsatellite loci, known as homoplasy. Only one strain, N. caninum Liverpool (Nc-Liv), has been genome sequenced and compared with its closest relative, Toxoplasma gondii (T. gondii). Hence, detailed population genomics studies based on whole-genome sequences from multiple strains worldwide are needed in order to better understand the current population genetic structure of Neospora, and ultimately to determine more effective vaccine candidates against bovine neosporosis. The aim of this review is to outline our current understanding of the molecular epidemiology and genomics of Neospora in juxtaposition with the closely related apicomplexan parasites Hammondia hammondi and T. gondii.

Molecular epidemiology and putative origin of hepatitis C virus in random volunteers from Argentina

AIM:To study the subtype prevalence and the phylogenetic relatedness of hepatitis C virus(HCV)sequences obtained from the Argentine general population,a large cohort of individuals was analyzed.METHODS:Healthy Argentinian volunteers(n=6251)from 12 provinces representing all geographical regions of the country were studied.All parents or legal guardians of individuals younger than 18 years provided informed written consent for participation.The corresponding written permission from all municipal authorities was obtained from each city or town where subjects were to be included.HCV RNA reverse transcription-polymerase chain reaction products were sequenced and phylogenetically analyzed.The 5’untranslated region(5’UTR)was used for RNA detection and initial genotype classification.The NS5B polymerase region,encompassing nt 8262-8610,was used for subtyping.RESULTS:An unexpectedly low prevalence of HCV infection in the general population(0.32%)was observed.Our data contrasted with previous studies that reported rates ranging from 1.5%to 2.5%,mainly performed in selected populations of blood donors or vulnerable groups.The latter values are in keeping with the prevalence reported by the 2007 Argentinian HCV Consensus(approximately 2%).HCV subtypes weredistributed as follows:1a(25%),1b(25%),2c(25%),3a(5%),and 2j(5%).Two isolates ascribed either to genotype 1(5%)or to genotype 3(5%)by 5’UTR phylogenetic analysis could not be subtyped.Subtype 1a sequences comprised a highly homogeneous population and clustered with United States sequences.Genotype1b sequences represented a heterogeneous population,suggesting that this genotype might have been introduced from different sources.Most subtype 2c sequences clustered close to the 2c reported from Italy and Southern France.CONCLUSION:HCV has a low prevalence of 0.32%in the studied general population of Argentina.The pattern of HCV introduction and transmission in Argentina appears to be a consequence of multiple events and different for each subtype.

Biochemical and Molecular Epidemiology of Cancer

Examples of practical approaches to molecular epidemiology of human cancer are described. Biomarkers of carcinogen exposure or inherited host factors for cancer susceptibility are discussed. Major advances have been made in the detection of carcinogen-macromolecular adducts through the use of high performance liquid chromatography, immunoaffinity chromatography, the 32P-postlabeling assay, enzyme immunoassays, gas chromatography/mass spectroscopy and synchronous spectrophotofluorimetry. The polycyclic aromatic hydrocarbon- DNA adducts are the most extensively studied in this field and together with antibodies to these adducts found in human serum, they have become useful indicators of exposure to carcinogens. Assays for various kinds of alky 1-DNA adducts have also been developed and the presence of these adducts have been documented in human tissues. Carcinogen-protein adducts have proven to be useful molecular dosimeters of carcinogen exposure. For example, 4-aminobiphenyl hemoglobin adducts are highly correlated with exposure to tobacco smoke. The study of the molecular aspects of interindividual differences in the metabolism and activation of xenobiotics and other genetic markers [DNA-restriction fragment length polymorphisms (RFLPs), mutations, and functional loss of specific genes in carcinogenesis] is an emerging new field that is discussed in the context of genetic susceptibility to cancer. The cytochrome P450 phenotypes and acetylation phenotype are examples of genetic markers that indicate an individual's potential for metabolism of exogenous substances. Further, inherited genetic polymorphic markers, e.g., DNA-RFLPs at protooncogene loci (HRAS-1 and L-myc) have been examined in a case-control study of lung cancer. Data concerning mutations of protooncogenes (H-, K-, and N-RAS) and tumor suppressor genes (retinoblastoma and p53 genes) in various common cancers are providing evidence of multiple genetic lesions that occur during the multistage process of carcinogenesis.

Molecular epidemiology of hepatitis B virus in Asia

Although safe and effective vaccines against hepatitis B virus(HBV) have been available for three decades, HBV infection remains the leading cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma(HCC) worldwide, especially in Asian countries. HBV has been classified into at least 9 genotypes according to the molecular evolutionary analysis of the genomic DNA sequence and shown to have a distinct geographical distribution. Novel HBV genotypes/subgenotypes have been reported, especially from Southeast Asian countries. The clinical characteristics and therapeutic effectiveness of interferon(IFN) and nucleos(t)ide analogues vary among different HBV genotypes. Mutations at T1653 C in subgenotype C2 from Japan and South Korea, C/A1753 T and C1858 T in subgenotype C1 from Vietnam, and C1638 T and T1753 V in subgenotype B3 from Indonesia were reported to be associated with advanced liver diseases including HCC. Genotype distribution in Japan has been changed by an increasing ratio of subgenotype A2 in chronic hepatitis B. While a large number of epidemiological and clinical studies have been reported from Asian countries, most of the studies were conducted in developed countries such as Taiwan, China, South Korea and Japan. In this review, the most recent publications on the geographical distribution of genetic variants of HBV and related issues such as disease progression and therapy in Asia are updated and summarized.

Scotomas in molecular virology and epidemiology of hepatitis C virus

In the 1970s,scientists learned of a new pathogen causing non-A,non-B hepatitis.Classical approaches were used to isolate and characterize this new pathogen,but it could be transmitted experimentally only to chimpanzees and progress was slow until the pathogen was identified as hepatitis C virus(HCV)in 1989.Since then,research and treatment of HCV have expanded with the development of modern biological medicine:HCV genome organization and polyprotein processing were delineated in 1993;the first three-dimensional structure of HCV nonstructural protein(NS3 serine protease)was revealed in 1996;an infectious clone of HCV complementary DNA was first constructed in 1997;interferon and ribavirin combination therapy was established in 1998 and the therapeutic strategy gradually optimized;the HCV replicon system was produced in1999;functional HCV pseudotyped viral particles were described in 2003;and recombinant infectious HCV in tissue culture was produced successfully in 2005.Recently,tremendous advances in HCV receptor discovery,understanding the HCV lifecycle,decryption of the HCV genome and proteins,as well as new anti-HCV compounds have been reported.Because HCV is difficult to isolate and culture,researchers have had to avail themselves to the best of modern biomedical technology;some of the major achievements in HCV research have not only advanced the understanding of HCV but also promoted knowledge of virology and cellular physiology.In this review,we summarize the advancements and remaining scotomas in the molecular virology and epidemiology of HCV.

Molecular epidemiology of DNA repair gene polymorphisms and head and neck cancer

Although tobacco and alcohol consumption are two common risk factors of head and neck cancer （HNC）, other specific etiologic causes, such as viral infection and genetic susceptibility factors, remain to be understood. Hu- man DNA is often damaged by numerous endogenous and exogenous mutagens or carcinogens, and genetic vari- ants in interaction with environmental exposure to these agents may explain interindividual differences in HNC risk. Single nucleotide polymorphisms （SNPs） in genes involved in the DNA damage-repair response are reported to be risk factors for various cancer types, including HNC. Here, we reviewed epidemiological studies that have assessed the associations between HNC risk and SNPs in DNA repair genes involved in base-excision repair, nucleotide-excision repair, mismatch repair, double-strand break repair and direct reversion repair pathways. We found, however, that only a few SNPs in DNA repair genes were found to be associated with significantly in- creased or decreased risk of HNC, and, in most cases, the effects were moderate, depending upon locus-locus in- teractions among the risk SNPs in the pathways. We believe that, in the presence of exposure, additional pathway- based analyses of DNA repair genes derived from genome-wide association studies （GWASs） in HNC are needed.

Molecular epidemiology of HIV-1 strains in the south-east and east of Turkey

Objective:To detect the subtype characterization and drug-resistant mutations in HIV-1 strains after the refugee movement from Syria to Turkey between 2011 and 2014 in south east border lines. Methods: A total of 65 patients were included in this study, of which 57(88%) patients were antiretroviral therapy-naive patients. HIV-1 RNA was detected and quantii ed by realtime PCR assay. HIV-1 subtypes and circulating recombinant forms(CRFs) were identii ed by phylogenetic analysis(neighbor-joining method), and drug-resistant mutations were analyzed.Results: Three major HIV groups were indicated. Two of these groups were located in subtype B. The other group showed heterogeneity. Subtype B(48/65, 73.8%), followed by CRFs(12/65, 18.5%) was the most common strain. Subtype of CRFs consisted of CRF01_AE(9/65, 13.8%) and CRF02_AG(3/65, 4.6%). Subtype C(1/65, 1.5%), sub-subtypes A1(2/65, 3.1%) and F1(2/65, 3.1%) were also detected with low prevalence. The rate of overall primary antiretroviral resistance was 4.9%(3/61). Drug-resistant rate for non-nucleoside reverse transcriptase inhibitors was 4.9%. The thymidine analogue mutation rate was 13.1%(8/61).Conclusions: HIV molecular epidemiology studies are necessary to determine transmission patterns and spread. Subtype B and CRF01_AE, CRF02_AG are the most prevalent strains in the south-east of Turkey. However, subtype C, sub-subtypes A1 and F1 are of low prevalence but persist in the south-east of Turkey. In the near future, changing of HIV epidemiology will be possible in Turkey due to migration movement in border lines and resistance testing will play an important role in HIV management.

Molecular Epidemiology and Risk Factors of Herpes Simplex Virus Type 2 in Pregnant Women

Background:Herpes simplex virus type 2(HSV-2)infection is the main cause of genital and neonatal herpes infections.It has considerable public health importance among women as the virus may lead to adverse outcomes in pregnancy and neonatal infection.This study determines the molecular epidemiology and risk factors ofHSV-2 infection among pregnant women.Methods:In this cross-sectional study,all pregnant women admitted to three university hospitals for natural birth and Caesarean sections were enrolled.HSV detection and typing were carried out based on PCR and reverse dot blotting method,respectively.ANOVA and bivariate correlations were used to analyze the data.Results:In this study,the prevalence of genital herpes infection was 5.7%.A significant positive correlation was found between age group<25 years and HSV-2 shedding(P=0.026).Twelve participants(60%)with HSV-2 shedding were younger than 25.A significant correlation was found between the presence of genital lesion and HSV-2(P=0.02).Among participants with HSV-2 infection,the use of condom was low.Neonatal complications were not seen in newborns from mothers with HSV-2 shedding.Conclusion:PCR assay may help in promoting early diagnosis and more effective treatment for patients.Also,it shortens hospital stay and enhances patients?condition.HSV-2 transmission is rapid following the onset of sexual activity and likely to result in the significant prevalence of genital disease.

Molecular Epidemiology of Nontuberculous Mycobacteria in a German CF Center and Clinical Course of NTM Positive Patients

Goal of this study was to analyse the clinical course of cystic fibrosis (CF) patients with nontuberculous mycobacteria (NTM) in their respiratory secretions and to investigate the molecular epidemiology of the most prevalent NTM species by multilocus sequence analysis (MLSA). The respiratory specimen and the clinical parameters forced expiratory volume in one second (FEV1), body-mass-index (BMI), erythrocyte sedimentation rate (ESR) 1 h and immunoglobulin G (IgG) of 357 CF patients, 0 - 52.4 years, mean FEV1 2009 81.5% pred were analysed between 1998 and 2010. In 13 patients NTM were detected. 12 of 13 patients carried M. abscessus, for one patient the NTM species was not characterized. 4 patients carried a second NTM species (M. avium, M. chelonae (2x), M. intracellulare). 6 patients exhibited a significant decline in FEV1, however changes in BMI, IgG and ESR were discordant. Molecular genotyping of M. abscessus isolates revealed a unique MLSA pattern in 6 patients. 2 patients harboured identical strains, and one patient a closely related strain. Whether the presence of identical strains is attributed to the acquisition of NTM clones from common environmental sources or to patient-to-patient transmission cannot be definitely clarified. Although cross-in- fection of the three patients with identical/closely related strains in the present cohort is highly unlikely, we recommend strict hygiene measures for all CF patients harbouring NTM.

Carcinoma of the stomach: A review of epidemiology, pathogenesis, molecular genetics and chemoprevention

Carcinoma of the stomach is still the second most common cause of cancer death worldwide, although the incidence and mortality have fallen dramatically over the last 50 years in many regions. The incidence of gastric cancer varies in different parts of the world and among various ethnic groups. Despite advances in diagnosis and treatment, the 5-year survival rate of stomach cancer is only 20 per cent. Stomach cancer can be classified into intestinal and diffuse types based on epidemiological and clinicopathological features. The etiology of gastric cancer is multifactorial and includes both dietary and nondietary factors. The major diet-related risk factors implicated in stomach cancer development include high content of nitrates and high salt intake. Accumulating evidence has implicated the role of Helicobacter pylori (H. pylori) infection in the pathogenesis of gastric cancer. The development of gastric cancer is a complex, multistep process involving multiple genetic and epigenetic alterations of oncogenes, tumor suppressor genes, DNA repair genes, cell cycle regulators, and signaling molecules. A plausible program for gastric cancer prevention involves intake of a balanced diet containing fruits and vegetables, improved sanitationand hygiene, screening and treatment of H. pylori infection, and follow-up of precancerous lesions. The fact that diet plays an important role in the etiology of gastric cancer offers scope for nutritional chemoprevention. Animal models have been extensively used to analyze the stepwise evolution of gastric carcinogenesis and to test dietary chemopreventive agents. Development of multitargeted preventive and therapeutic strategies for gastric cancer is a major challenge for the future.

Epidemiology and molecular genotyping of echinostome metacercariae in Filopaludina snails in Lamphun Province,Thailand

Objective:To analyze the prevalence of echinostome metacercariae in Filopaludina dorliaris(F.dorliaris)and Filopaludina martensi martensi(F.martensi martensi)and genotype variation of echinostome metacercariae by using random amplified polymorphic DNA(RAPD)analysis.Methods:Filopaludina sp.snails were collected from eight localities of Lamphun Province,Northern Thailand and examined for echinostome metacercariae.RAPD-PCR was used lo analyze genotype variation of echinostome metacercariae.Results:A total of 3 226 F.dorliaris and F.martensi martensi snails were collected from eight localities.The overall prevalences of echinostome metacercariae in F.dorliaris and F.martensi martensi were 40.89%and 36.27%,while the intensity of infection was 20.37 and 12.04,respectively.The dendrogram constructed base on RAPD profiles,4 well supported domains were generated;(i)group of metacercariae from Ban Hong,Mae Ta,Meaung,Pa Sang,Toong Hua Chang,and Weang Nong that were clustered in the group of E.revolutum,(ii)Ban Thi,(iii)Lee,and(iv)3 adults of an out group.Condusioiis:This research demonstrated RAPD profiling has been a useful tool to detect DNA polymorphisms to determine genetic relationship between echinostome metacercariae in Lamphun Province,Northern Thailand.

Molecular Epidemiology and Clinical Features of Haemophilus influenzae among Hospitalized Children with Community-acquired Pneumonia in Chengde,China

Community-acquired pneumonia(CAP)is an acute lung infection that is caused by several different pathogens and is associated with significant morbidity and mortality.The high global incidence of CAP poses a heavy disease and economic burden to patients,especially children.Respiratory illnesses such as pneumonia and influenza are the fourth leading cause of death in China[1].The top 3 etiologic pathogens of CAP in the Asia-Pacific region are Streptococcus pneumoniae,Haemophilus influenzae(H.influenzae),and Mycoplasma pneumoniae(M.pneumoniae).

Molecular Epidemiology of Klebsiella pneumoniae from Clinical Bovine Mastitis in Northern Area of China,2018–2019

Klebsiella pneumoniae(K.pneumonia,KpⅠ)is a predominate inducement of bovine mastitis,which is associated with high mortality and milk yield reduction.However,data is lacking on the molecular characteristics of bovine K.pneumoniae,limiting the risk assessment of its transmission through the food chain.Herein,we investigated the prevalence of K.pneumoniae in 6301 clinical mastitis(CM)milk samples from dairy cattle in northern area of China.In total,183 K.pneumoniae isolates were recovered,with detection rates of 3.0% and 2.8% in 2018 and 2019,respectively.Like human clinical K.pneumoniae,all CM K.pneumoniae isolates belonged to one of three phylogroups:KpⅠ(n=143),Klebsiella.quasipneumoniae subsp.similipneumoniae(KpⅡ-B)(n=37),and Klebsiella variicola(KpⅢ)(n=3).We detected the extendedspectrum β-lactamase-encoding genes bla_(SHV-2a),blac_(CTX-M-14),and bla_(CTX-M-15),as well as clpC,lpfA,lacI,lacZ,lacY,and the fecABDEIR operon in the KpⅠ isolates,which may contribute to their pathogenicity and host adaptability in cows.The high prevalence of KpⅠ in dairy farms may be problematic,as it showed relatively higher rates of antibiotic resistance and virulence gene carriage than the KpⅡ-B and KpⅢ isolates.Furthermore,we observed distinct differences in population structure between CM-and human infection-associated KpⅠ isolates,with the genes associated with invasive infection in humans rarely being observed in bovine isolates,indicating that few CM-associated K.pneumoniae isolates pose a threat to human health.Nevertheless,bovine KpⅡ-B isolates shared a high level of nucleotide sequence identity with isolates from human infections and frequently carried the nitrogen-fixation gene nif,suggesting an association between KpⅡ-B isolates from cattle and humans,and plant-derived bacteria.