Infection In Dairy Cattle Research Articles

Molecular characterization of extended-spectrum β-lactamase-producing Escherichia coli isolated from postpartum uterine infection in dairy cattle in India.

Background and Aim:Selection and dissemination of plasmid-encoded extended-spectrum β-lactamase (ESBL) among Enterobacteriaceae confers resistance to beta-lactam antibiotics. The purpose of this study was to determine the prevalence and molecular characteristics of ESBL-producing organisms isolated from dairy cattle with a uterine infection.Materials and Methods:Bacterial isolates (n=62) were characterized by biochemical test for genus and species determination. Antimicrobial susceptibility tests were performed by Kirby–Bauer disk diffusion method using panel of antibiotics for initial screening of ESBL organism. Phenotypic confirmation of ESBL-suspected strains was done by combination disk method and double-disk method. Multiplex polymerase chain reaction (PCR) was carried out for phylogrouping of Escherichia coli isolates as well as for genotyping ESBL genes. Enterobacterial repetitive intergenic consensus-PCR method was used for genotypic characterization of isolates.Results:Antibiotic susceptibility profile of E. coli (n=40) isolates showed high rates of resistance for ampicillin (95.0%), cefpodoxime (97.5%), cefotaxime (87.5%), and ceftriaxone (70%). However, low rates of resistance were observed for cefoxitin (25%), amoxicillin/clavulanic acid (20%), ceftazidime (17.5%), gentamicin (10%), and ertapenem (7.5%). A total of 39/40 E. coli isolates were confirmed as ESBL with Epsilometer test as well as the genotypic method and 28 (70%) of them were multidrug-resistant. Genotype blaCTX-M was observed as a predominant beta-lactamase type with the preponderance of CTX-M Group 1. The following combinations were observed: blaTEM+ blaCTX-M in 15 (36.2%) isolates, blaTEM/blaSHV in 8 (5.2%) isolates, and blaCTX-M/blaSHV in 6 (5.2%) isolates. The phylogenetic grouping of E. coli strains revealed the highest prevalence for B1 (22.0%) followed by A (20%).Conclusion:This report shows a high frequency of ESBL E. coli from cattle with postpartum uterine infections. These isolates showed reduced susceptibility to common antibiotics used for the treatment of uterine infections greater affecting the therapeutic outcome.

Current Developments in the Epidemiology and Control of Enzootic Bovine Leukosis as Caused by Bovine Leukemia Virus.

Enzootic Bovine Leukosis (EBL) caused by the bovine leukemia virus (BLV) has been eradicated in over 20 countries. In contrast, the U.S. and many other nations are experiencing increasing prevalence in the absence of efforts to control transmission. Recent studies have shown that BLV infection in dairy cattle has a greater impact beyond the long-recognized lymphoma development that occurs in <5% of infected cattle. Like other retroviruses, BLV appears to cause multiple immune system disruptions, affecting both cellular and humoral immunity, which are likely responsible for increasingly documented associations with decreased dairy production and decreased productive lifespan. Realization of these economic losses has increased interest in controlling BLV using technology that was unavailable decades ago, when many nations eradicated BLV via traditional antibody testing and slaughter methods. This traditional control is not economically feasible for many nations where the average herd antibody prevalence is rapidly approaching 50%. The ELISA screening of cattle with follow-up testing via qPCR for proviral load helps prioritize the most infectious cattle for segregation or culling. The efficacy of this approach has been demonstrated in at least four herds. Breeding cattle for resistance to BLV disease progression also appears to hold promise, and several laboratories are working on BLV vaccines. There are many research priorities for a wide variety of disciplines, especially including the need to investigate the reports linking BLV and human breast cancer.

Isolation and Antimicrobial Susceptibility Profile of Escherichia coli O157 : H7 from Raw Milk of Dairy Cattle in Holeta District, Central Ethiopia

A cross-sectional study was conducted in small, medium, and large-scale dairy farms of Holeta district to isolate, identify, and antimicrobial susceptibility profile of Escherichia coli O157 : H7 in raw milk of dairy cattle. A total of 210 lactating cows were selected for raw milk samples, and 19% (40/210) were found to be positive for E. coli whereas 5.2% (11/210) were confirmed as E. coli O157 : H7 positive using the Escherichia coli O157 latex test. Accordingly, all E. coli was highly susceptible to Ciprofloxacin (100%), Gentamycin (100%), Oxytetracycline (100%), and Tetracycline (63.63%). Furthermore, the resistance of 72.73%, 54.54%, 54.54%, and 45.45% was developed to Cefoxitin, Sulphamethoxazole, Cloxacillin, and Streptomycin, respectively. Factors such as parity, age, body condition, herd size, milk yield, udder hygiene, and udder lesion showed a statistically significant ( p &lt; 0.05 ) association with the occurrence of E. coli infection in dairy cattle. In conclusion, in this study, a higher prevalence of Escherichia coli O157 : H7 and its drug susceptibility profile is an alarm for the health of the public, and awareness creation to the farm owners and the community is recommended.

Genetic recombination of bovine viral diarrhea virus subgenotype -1a and -1c in persistently infected dairy cattle

The bovine viral diarrhea virus (BVDV) is a major viral pathogen in cattle worldwide. In Indonesia, diversity in subgenotypes of BVDV-1 has been observed, with the highest proportion of subgenotype -1a, followed by -1c, -1b, and -1d. So far, phylogenetic analysis of BVDV-1 is based on nucleotide sequences of the 5′ UTR and partial NS5B regions. Accuracy in identifying the subgenotype and antigenic type is critical for vaccine development and effective vaccination. The aim of this study was to determine genetic recombination of BVDV through phylogenetic analysis of five different regions (5′ UTR, NPro, E2, NS3, and NS5B) of BVDV in persistently infected dairy cattle. Five isolates were sequenced using next-generation sequencing, and data were analyzed with the CLC Genomic Workbench 9.0 and MEGA-X programs. Phylogenetic analysis based on the 5′ UTR (275 nt), NPro (504 nt), E2 (1,122 nt), NS3 (2,049 nt), and NS5B (2,157 nt) regions indicated that one BVDV isolate from Banyumas, Central Java, could be classified into different subgenotypes based on the E2 region (-1c), but the same subgenotype based on the other four regions (-1a), suggesting the presence of genetic recombination of the BVDV subgenotypes -1a and -1c in persistently infected dairy cattle.

High quality genome sequence of Treponema phagedenis KS1 isolated from bovine digital dermatitis

Treponema phagedenis KS1, a fastidious anaerobe, was isolated from a bovine digital dermatitis (BDD)-infected dairy cattle in Chungnam, Korea. Initial data indicated that T. phagedenis KS1 exhibited putative virulent phenotypic characteristics. This study reports the whole genome assembly and annotation of T. phagedenis KS1 (KCTC14157BP) to assist in the identification of putative pathogenicity related factors. The whole genome of T. phagedenis KS1 was sequenced using PacBio RSII and Illumina HiSeqXTen platforms. The assembled T. phagedenis KS1 genome comprises 16 contigs with a total size of 3,769,422 bp and an overall guanine-cytosine (GC) content of 40.03%. Annotation revealed 3,460 protein-coding genes, as well as 49 transfer RNA- and 6 ribosomal RNA-coding genes. The results of this study provide insight into the pathogenicity of T. phagedenis KS1.

Genotypic and Phenotypic Characterization of Treponema phagedenis from Bovine Digital Dermatitis.

This study aimed to isolate and characterize Treponema spp. from bovine digital dermatitis (BDD)-infected dairy cattle. Seven isolates were characterized in this study. Isolates exhibited slow growth, and colonies penetrated the agar and exhibited weak β-hemolysis. Round bodies were observed in old and antibiotic-treated cultures. Cells ranged from 9–12 µm in length, 0.2–2.5 µm in width, and were moderately spiraled. The 16S rRNA analysis revealed the isolates as Treponema phagedenis with >99% sequence homology. Isolates had alkaline phosphatase, acid phosphatase, β-galactosidase, N-acetyl-β-glucosaminidase, esterase (C4), esterase lipase (C8), naphthol-AS-BI-phosphohydrolase, and β-glucuronidase activities. Low concentrations of ampicillin, erythromycin, and tetracycline were required to inhibit the growth of isolates. Formic, acetic, and butyric acids were produced, while propionic acid was significantly utilized, indicating its essentiality for treponemal growth. The isolates shared the same characteristics and, therefore, were considered as a single strain. Isolate HNL4 was deposited as a representative isolate (Treponema phagedenis KS1). The average nucleotide identity of strain KS1 showed a small difference with the human strain (99.14%) compared with bovine strain (99.72%). This study was the first to isolate and characterize Treponema phagedenis from BDD in Korea and, hence, it delivered pathogenicity-related insights and provided valuable information that can be used for the management of BDD.

Risk factor analysis of bovine leukemia virus infection in dairy cattle in Egypt

Identification of the risk factors associated with Enzootic bovine leukosis (EBL) is essential for the adoption of potentially prevention strategies. Accordingly, our objectives were to determine the geographic distribution of Bovine Leukemia Virus (BLV) infection and identify the risk factors associated with cow-level BLV infection in the Egyptian dairy cattle. A cross-sectional study was conducted on 1299 mixed breed cows distributed over four provinces in the Nile Delta of Egypt in 2018. The randomly selected cows on each farm were serologically tested for BLV, and the cow’s information was obtained from the farm records. Four variables (geographic location, herd size, number of parities, and age) were used for risk analysis. A total of 230 serum samples (17.7 %) were serologically positive for BLV. The highest prevalence of BLV infection was associated with parity (OR = 3.4, 95 %CI 2.4–4.9) with 80 % probability of being BLV-positive at parity ≥5, followed by herd size (OR = 1.8, 95 %CI 1.4–2.2). However, geographic location seems to have no impact on the prevalence of BLV infection in Egypt. Our findings strongly indicate that the intensive surveillance and effective prevention strategies against BLV infection in Egypt should be provided to multiparous cows with ≥5 parities and live in large farm with more than 200 cows.

Salmonella enterica Subsp. Enterica Serovar Reading Infection in Dairy Cattle and Buffaloes Suffering from Chronic Diarrhea

The study was aimed at dairy cattle and buffaloes presented with the history of chronic diarrhea to isolate and detect Salmonella spp. Fecal samples were collected directly from the rectum of diarrheic cattle. Isolation and identification of the microorganisms was performed and confirmed on the basis of their morphology, staining, cultural, biochemical tests and sero-typing. Salmonella positive animals presented variable degree and frequency of diarrhea. Salmonellosis was confirmed in seven per cent of cattle and buffaloes suffering from chronic diarrhea. The serotype detected in the 7 Salmonella strains was Salmonella enterica subsp. enterica serovar Reading. Salmonella isolates recovered from dairy cattle had relatively variable resistance to various antimicrobial agents. The antimicrobial susceptibility pattern of the isolates showed isolates were resistant to amoxicillin, tetracycline, ampicillin, ceftriaxone. Whereas 87.5 percent, 75.0 per cent and 62.5 percent susceptible to ciprofloxacin, gentamicin and co-trimoxazole respectively. This study provides updated information on the bio-incidence and susceptibility patterns of Salmonella in dairy animals suffering chronic diarrhea. Isolation and serotyping of Salmonella enterica subsp. enterica serovar Reading is new to the study and contribute to our understanding that there is shift in increases in susceptibility of dairy herds to acquire new strains.

Bayesian latent class estimation of sensitivity and specificity parameters of the PMS-PCR test for the diagnosis of cattle sub-clinically infected with Mycobacterium avium subsp. paratuberculosis

The objective of this study was to estimate the performance of the peptide magnetic separation PCR test (PMS-PCR) for the diagnosis of Mycobacterium avium subsp. paratuberculosis (MAP) in sub-clinically infected dairy cattle. Twenty-one herds were randomly selected from a source population of 131 commercial dairy herds with a known history of MAP infection, located in the De Los Rios and De Los Lagos regions, in southern Chile. In the selected herds, all milking cows with ≥2 parities and without any clinical signs were sampled, collecting feces and blood-serum samples. The PMS-PCR test was used to analyze the fecal samples, while serum samples were analyzed using a commercial ELISA kit. A Bayesian latent class model was used to estimate the sensitivity (Se) and specificity (Sp) of the diagnostic tests.A total of 1381 animals were sampled in the 21 selected dairy herds, with an average sample size of 65 animals per herd (range 10–721). The PMS-PCR test had a greater Se than the ELISA test, with a median of 85.5 % (posterior probability interval (PPI) 95 %: 79.3–91.0%), while the ELISA test presented a median of 21.7 % (95 % PPI: 18.3–25.4%). On the other hand, the ELISA test had a better Sp than the PMS-PCR test, with a median of 97.7 % (95 % PPI: 96.6–98.5%), whereas PMS-PCR presented a median of 90.8 % (95 % PPI: 88.3–93.9%). Model results showed that PMS-PCR has a better Se than all available tests for MAP diagnosis in subclinical animals. However, this test should be used with care in herds with high infection rates, where a high MAP environmental load is expected, potentially increasing the frequency of false positive cases due to the pass-through phenomenon.

Serological and molecular evidence of Mycobacterium bovis in dairy cattle and dairy farm workers under the intensive dairy production system in Egypt.

We employed a combination of diagnostic tests including single cervical tuberculin test (SCT), rapid lateral-flow test (RT), TB-Feron, conventional PCR and culture to investigate the prevalence of Mycobacterium bovis infections in dairy cattle under the intensive dairy production system in Egypt. In total, 2710 dairy cows from 11 herds in six Governorates were tested by SCT and 444 (16·4%) were TB reactors. Only 65 cows responded to M. bovis antigen by RT and TB-Feron. A postmortem examination showed that 49 (75·4%) of slaughtered cows have visible lesions. Testing of 215 out 444 SCT reactor cows by culture and PCR using blood and milk samples revealed that M. bovis were more frequently identified in the blood (6·1-20·5%) than milk (2·3-5·6%) samples. Additionally, in this study, we investigated the prevalence and potential risk factors associated with M. bovis infection in dairy farm workers. Overall, 100 dairy farm workers were tested using QuantiFERON-TB Gold In-Tube test to detect Mycobacterium tuberculosis complex (MTC) and 35 (35%) were positive. In all, 23 (23%) of MTC positive were M. bovis positive using PCR. Mycobacterium bovis positive cases were associated with workers who had respiratory signs and did not wash or disinfect their hands after handling cows. The prevalence of M. bovis in dairy cows and dairy farm workers under the intensive dairy production system in Egypt is high. It is therefore essential to disseminate effective prevention and control measures to prevent the spread of M. bovis between dairy cows and dairy workers. This study revealed that the use of RT or TB-Feron as an ancillary test of SCT reactor cows resulted in a significant reduction in the SCT false-positive slaughtered cows. A high prevalence of M. bovis infection among farm workers provides evidence of occupational risk in the intensive dairy production system in Egypt.

Staphylococcus aureus persistence properties associated with bovine mastitis and alternative therapeutic modalities.

Staphylococcus aureus is an important agent of contagious bovine intramammary infections in dairy cattle. Its ability to persist inside the udder is based on the presence of important mechanisms such as its ability to form biofilms, polysaccharide capsules small colony variants, and their ability to invade professional and nonprofessional cells, which will protect S. aureus from the innate and adaptive immune response of the cow, and from antibiotics that are no longer considered to be sufficient against S. aureus bovine mastitis. In this review, we present the recent research outlining S. aureus persistence properties inside the mammary gland, including its regulation mechanisms, and we highlight alternative therapeutic strategies that were tested against S. aureus isolated from bovine mastitis such as the use of probiotic bacteria, bacteriocins and bacteriophages. Overall, the persistence of S. aureus inside the mammary gland remains a pressing veterinary problem. A thorough understanding of staphylococcal persistence mechanisms will elucidate novel ways that can help in the identification of novel treatments.

Molecular detection of Anaplasma spp. in dairy cattle in southern Xinjiang, China.

Bovine anaplasmosis is caused by a group of obligate intracellular bacteria belonging to the genus Anaplasma, which are transmitted by ticks. This study was conducted to determine the prevalences and molecular characterization of Anaplasma spp. in dairy cattle in the upper reaches of the Tarim River in Xinjiang, China. Using polymerase chain reaction (PCR) and sequencing approaches, DNA of Anaplasma spp. was detected in 16 of 493 (3.2%) blood samples from dairy cattle. Positive rates were 0.2% (1/493), 0.4% (2/493), 0.2% (1/493), 2.4% (12/493) and 2.4% (12/493) for A. bovis, A. ovis, A. phagocytophilum like strain, A. phagocytophilum and A. platys like strain, respectively. Anaplasma phagocytophilum and A. platys like strain co-infection was detected in 12 samples. To our knowledge, this is the first report of A. ovis infection in dairy cattle in Xinjiang. This study provides new data on the prevalences of Anaplasma spp. in cattle in Xinjiang, which will help to formulate appropriate control strategies for these pathogens in this area.

Bartonella bovis and Bartonella chomelii infection in dairy cattle and their ectoparasites in Algeria

Bartonella are blood-borne and vector-transmitted bacteria, some of which are zoonotic. B. bovis and B. chomelii have been reported in cattle. However, no information has yet been provided on Bartonella infection in cattle in Algeria. Therefore, 313 cattle from 45 dairy farms were surveyed in Kabylia, Algeria, in order to identify Bartonella species infecting cattle using serological and molecular tests. In addition, 277 ticks and 33 Hippoboscidae flies were collected. Bartonella bovis and B. chomelii were identified as the two species infecting cattle. Bartonella DNA was also amplified from 6.8 % (n = 19) of ticks and 78.8 % (n = 26) of flies. Prevalence of B. bovis DNA in dairy cattle was associated both with age and altitude. This study is the first one to report of bovine bartonellosis in Algeria, both in dairy cattle and in potential Bartonella vectors, with the detection of B. bovis DNA in tick samples and B. chomelii in fly samples.

Lack of association between amino acid sequences of the bovine leukemia virus envelope and varying stages of infection in dairy cattle

We collected 724 blood samples from dairy cattle from six Mexican states, and tested them for the presence of antibodies against BLV using a commercial ELISA test. Our study groups consisted of 32 samples: 12 asymptomatic cows, 12 cows with lymphocytosis and 8 samples of tumor tissue of the abomasum and heart of cattle with lymphoma. We designed three pairs of primers to amplify the complete BLV env gene, and obtained a fragment of 1548 nucleotides in length with the sequenced products. According to the phylogenetic tree we constructed to identify the viral genotype, 96.87 % of the sequences grouped into genotype 1, while a single sample from a cow with lymphocytosis (3.13 %) was associated with genotype 3 sequences. The similarity between the Mexican BLV sequences ranged from 0.985–1.00. In addition, the proportion of non-synonymous and synonymous mutations indicated negative selection. We did not identify any conserved residues in the viral protein sequences that could be related to BLV infection stage in cattle. Proviral quantification was performed using quantitative polymerase chain reaction, and we used Mood´s median test as statistical analysis. We found no significant association between proviral load and phase of infection. The sequences showed high similarity without any association between BLV surface glycoprotein and the different infection stages, nor differences in the proviral load. BLV genotype 1 was identified as prevalent in the studied samples, and for the first time in Mexico, we identified BLV genotype 3 in cattle.

Determination of the prevalence and intensity of Fasciola hepatica infection in dairy cattle from six irrigation regions of Victoria, South-eastern Australia, further identifying significant triclabendazole resistance on three properties

Fasciola hepatica (liver fluke) is a widespread parasite infection of livestock in Victoria, South-eastern Australia, where high rainfall and a mild climate is suitable for the main intermediate host Austropeplea tomentosa. The aims of this study were to quantify the prevalence and intensity of F. hepatica in dairy cattle in the irrigated dairy regions of Victoria and determine if triclabendazole resistance was present in infected herds. Cattle in 83 herds from the following six irrigation regions were tested for F. hepatica: Macalister Irrigation District (MID), Upper Murray (UM), Murray Valley (MV), Central Goulburn (CG), Torrumbarry (TIA) and Loddon Valley (LV). Twenty cattle from each herd were tested using the F. hepatica faecal egg count (FEC) as well as the coproantigen ELISA (cELISA). The mean individual animal true prevalence of F. hepatica across all regions was 39 % (95 % credible interval [CrI] 27%–51%) by FEC and 39 % (95 % CrI 27%–50%) by cELISA with the highest true prevalence (75–80 %) found in the MID. Our results show that 46 % of the herds that took part in this study were likely to experience fluke-associated production losses, based on observations that herd productivity is impaired when the true within-herd prevalence is > 25 %. Using the FEC and cELISA reduction tests, triclabendazole resistance was assessed on 3 herds in total (2 from the 83 in the study; and 1 separate herd that did not take part in the prevalence study) and resistance was confirmed in all 3 herds. This study has confirmed that F. hepatica is endemic in several dairy regions in Victoria: triclabendazole resistance may be contributing to the high prevalence in some herds. From our analysis, we estimate that the state-wide economic loss associated with fasciolosis is in the order of AUD 129 million (range AUD 38–193 million) per year or about AUD 50,000 (range AUD 15,000–75,000) per herd per year.

Prevalence and genetic diversity of bovine viral diarrhea virus in dairy herds of China

To determine the nationwide prevalence and genetic diversity of bovine viral diarrhea virus (BVDV) in China, 92 dairy farms with more than 500 animals in 19 provinces of China were surveyed in 2017. At each farm, ear notch samples from calves less than six months old and bulk tank milk (BTM) samples were collected. A total of 901 ear notch samples and 329 BTM samples from 183 tanks were sampled. A total of 20 (20/901, 2.22 %) ear notch samples from 10 (10/92, 10.86 %) farms tested positive for BVDV by IDEXX Antigen Point-of-Care (POC) Test kit and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). In addition, 80 of 183 (80/183, 43.7 %) BTM samples from 43 (43/92, 46.7 %) farms were identified as positive by qRT-PCR. The RNA of positive and suspect samples identified by qRT-PCR was subjected to 5′- untranslated region (UTR) amplification by nested RT-PCR and then sequenced. A total of 119 sequences were obtained and phylogenetic analysis of these 5′-UTR sequences revealed the presence of eight different subgenotypes of BVDV-1 including 1a (n = 37, 31.09 %), 1b (n = 5, 4.20 %), 1c (n = 34, 28.57 %), 1d (n = 2, 1.68 %), 1m (n = 25, 21.01 %), 1q (n = 6, 5.04 %), and two unknown subgenotypes which were tentatively typed as “BVDV-1v” (n = 8, 6.72 %) and “BVDV-1w” (n = 2, 1.68 %), respectively. BVDV-1a, 1c, and 1m were the dominant strains, collectively accounting for 80.67 % (96/119) of all sequences. Phylogenetic analysis based on selected N-terminal autoprotease (Npro) sequences confirmed the classification of the 5′-UTR sequences. In conclusion, the prevalence of BVDV persistent infection in dairy cattle was high and genetic diversity was high and increasing, revealing a serious threat to the health of cattle in China and highlighting the need for BVDV control.

Bovine besnoitiosis in an endemically infected dairy cattle herd in Italy: serological and clinical observations, risk factors, and effects on reproductive and productive performances.

Bovine besnoitiosis (Besnoitia besnoiti) is an emerging parasitic disease of cattle in Europe. This study reports a case of bovine besnoitiosis in a dairy farm housing 217 cattle in Italy. A serological screening was performed on the whole herd using the recommended approach of ELISA and confirmatory Western Blot. Seropositive animals were clinically examined to reveal symptoms and lesions of besnoitiosis. Risk factors and the effects of the parasite infection on reproductive and productive performances were evaluated. Histopathology and molecular analyses on tissues from a slaughtered cow affected by the chronic phase of the disease were carried out. An overall seroprevalence of 23.5%, which increased up to 43.5% considering only cows, was recorded. Clinical examination of 33 of the seropositive cows evidenced the presence of tissue cysts in at least one of the typical localizations (sclera, vulva, or skin) in 25 animals. Statistical analysis did not evidence any significative impact of the parasite infection on herd efficiency; however, a decrease of productive parameters was recorded in cows showing cutaneous cysts. Concerning the chronically affected cow, histopathology revealed B. besnoiti tissue cysts in the skin of the neck, rump, hind legs, eyelid and vulva, in the muzzle, in mucosal membranes of the upper respiratory tract, and in the lungs. Parasite DNA was detected also in masseter muscles, tonsils, mediastinal lymph nodes, liver, cardiac muscle, aorta wall, ovaries, uterus, and vulva. Bovine besnoitiosis continues to spread in the Italian cattle population. Breeders and veterinarians should be aware of this parasitic disease, and control programs should be developed based on surveillance through a diagnostic procedure including both clinical examination and laboratory tests.

Transmission of Coxiella burnetii to Calves from Infected Cows

Background:Coxiella burnetii is the causative agent of a very important disease with zoonotic potential. Infected cows represent risk for spreading of infection to humans and to other animals on farm and also to their offspring. There is possibility for calves from infected cows to be infected nearly after parturition or during intrauterine life. Studies have shown that Coxiella burnetii initially infects the placenta and subsequent spread to the fetus may occur either by haematogenous or by the amniotic-oral route providing congenital infection. The main objective of the present study is to determine the presence of Coxiella burnetii genome in milk serum of infected cows and blood serum of calves. Materials, Methods &amp; Results: A total of 200 blood serums from dairy cows were tested for presence of antibodies to Coxiella burnetii and nine of those were found positive. These animals compiled experimental group. From animals in experimental group milk samples during lactation, pregnancy and the postpartum period were collected. Samples were used for performing PCR test for determination of Coxiella burnetii presence in milk serum. On calving of each cow blood samples were taken from calves during first 24 hours after calving, from jugular vein. These blood samples were also used for PCR test to determine the presence of Coxiella burnetii. Milk serum analysis showed presence of Coxiella burnetii genome in serum, indicating on intermittent excretion. During lactation, the excretion of bacteria was greatest in the second stage when 80% of milk serum samples were positive for Coxiella burnetii. In the colostrums stage, there was a high percentage of Coxiella burnetii excretion through milk (50% of positive milk serum samples). The lowest percentage of excretion through milk was in the first stage of lactation. Analyzing blood serum samples from calves, taken on first day at calving using PCR method, all serums were positive for presence of Coxiella burnetii genome. Discussion:In animals, Coxiella burnetii is found in the reproductive system, both uterus and mammary glands, and may cause abortion or infertility. The high prevalence of Coxiella burnetii infection in dairy cattle with reproductive problems showed that these infected cattle play an important role in maintaining the infection and in disseminating of pathogenic agent to environment. The lowest percentage of excretion of Coxiella burnetii was in the first stage of lactation, amounting to 16.6%. With the transition to the next stage of lactation, a striking increase in the percentage of excretion was noticed. In the second stage it was 80%. In the third stage there was decrease to 40.6%. In colostrums stage percentage of excretion through milk was 50%. Similar results claims that excretion of Coxiella burnetii through milk starts after eight to twelve weeks of lactation in most cows. This period coincides with second stage of lactation as we divide it. Blood serums taken from calves were proven positive on Coxiella burnetii which indicates on intrauterine infection as described in the literature. Intrauterine infection takes place after placenta infection when bacteria penetrate the placenta and contaminate the amniotic fluid and gets aspirated or swallowed by fetus. Besides this,haematogenous spread can also occur.

Within-host evolution of bovine Staphylococcus aureus selects for a SigB-deficient pathotype characterized by reduced virulence but enhanced proteolytic activity and biofilm formation

Staphylococcus aureus is a major cause of bovine mastitis, commonly leading to long-lasting, persistent and recurrent infections. Thereby, S. aureus constantly refines and permanently adapts to the bovine udder environment. In this work, we followed S. aureus within-host adaptation over the course of three months in a naturally infected dairy cattle with chronic, subclinical mastitis. Whole genome sequence analysis revealed a complete replacement of the initial predominant variant by another isogenic variant. We report for the first time within-host evolution towards a sigma factor SigB-deficient pathotype in S. aureus bovine mastitis, associated with a single nucleotide polymorphism in rsbU (G368A → G122D), a contributor to SigB-functionality. The emerged SigB-deficient pathotype exhibits a substantial shift to new phenotypic traits comprising strong proteolytic activity and poly-N-acetylglucosamine (PNAG)-based biofilm production. This possibly unlocks new nutritional resources and promotes immune evasion, presumably facilitating extracellular persistence within the host. Moreover, we observed an adaptation towards attenuated virulence using a mouse infection model. This study extends the role of sigma factor SigB in S. aureus pathogenesis, so far described to be required for intracellular persistence during chronic infections. Our findings suggest that S. aureus SigB-deficiency is an alternative mechanism for persistence and underpin the clinical relevance of staphylococcal SigB-deficient variants which are consistently isolated during human chronic infections.

Detection of Bovine Leukemia Virus RNA in Blood Samples of Naturally Infected Dairy Cattle.

The viral expression in vivo, in bovine leukemia virus (BLV)-infected cattle, is considered to be restricted to extremely low levels, and the mitosis of infected B lymphocytes is regarded as the main mode of virus persistence within the infected host. In this study, the presence of BLV RNA in whole blood from seven asymptomatic cows naturally infected with BLV during one year, including a complete milking cycle and two delivery time points, was investigated by nested-PCR using the oligonucleotides complementary to the tax and pol gene. BLV RNA was detected in four cows at different time points, especially in high blood proviral load cows and around delivery time. This study describes for the first time the detection of free BLV RNA in blood from BLV-infected asymptomatic cows. The results obtained suggest the occurrence of persistent low-level expression of the tax and pol genes that could be a result of viral reactivation, within the asymptomatic period. This finding may be important in the pathogenesis of BLV infection, associated with the delivery period.