Respiratory Tract Of Cattle Research Articles

First isolation and characterization of Bovine Herpesvirus 1.2b (BoHV-1.2b ) strain from respiratory tract of cattle in Turkey

Restriksiyon Parça Uzunluk Polimorfizmi (RFLP) tekniği kullanılarak tüm genomun analizinde Bovine Herpesviruses-1 (BoHV-1) suşlarını BoHV-1.1, 1.2a, 1.2b olmak üzere üç alt tipte sınıflandırılmaktadır. Bu alt tipler klinik bulgular göre de değerlendirilmektedir ancak bu yönde henüz net bir ilişkilendirme yapılmamıştır. Çoğunlukla BoHV-1.2b, genital hastalık bulgularıyla ilişkilendirilmektedir. Bu çalışmada solunum yolu hastalığından alınan nazal sürüntü örneğinde BoHV-1 izolatı elde edilmiştir. Bu çalışmada ayrıca, genomun UL44 (Glikoprotein C) bölgesini hedefleyen filogenetik analiz ve UL39 - US3 bölgelerini hedefleyen, yeni geliştirilen bir multipleks PCR analizini takiben Hind III enzimi kullanılarak uygulanan RFLP ile alt tiplendirme gerçekleştirilmiştir. Aynı zamanda izolasyon başarısı iki farklı devamlı hücre hattında karşılaştırılmıştır. SFT-R hücre hattı, BoHV-1 saha örneklerinin izolasyonunda MDBK hücre hattına nazaran daha duyarlı bulunmuştur. Elde edilen izolat (ID:8640), BoHV-1.2b olarak sınıflandırılırken, Cooper suşundan serolojik olarak ayırt edilemediği değerlendirildi. Mevcut çalışma, BoHV-1.2b'nin Türkiye'deki ilk izolasyonunu bildirmesinin yanı sıra klinik solunum yolu hastalığından nadir tespit edilen BoHV-1.2b izolasyonunu bildirmektedir. Sonuçlar ayrıca kolay ve hızlı alt tipleme için PCR tabanlı RFLP analizinin etkinliğini gösteren veriler içermektedir. Ancak BoHV-1 saha izolatlarının genetik çeşitliliğine bağlı olarak bu teknik üzerine daha fazla araştırma yapılmasına ihtiyaç duyulmaktadır.

Identification and Genetic Characterization of Mycoplasma Species Affecting Respiratory System in Egyptian Cattle

Abstract: Mycoplasma species are the main important causative agents causing pneumonia in cattle, there are about 200 species or more. The current work aimed to investigate various mycoplasma spp. isolated from the respiratory tract of cattle by microbial culture, conventional polymerase chain reaction technique, and target gene sequencing with phylogenetic analysis. An initial screening was done to confirm the presence of mycoplasma spp. by culture on PPLO’s agar, digitonin sensitivity and biochemical tests. 129 isolates were characterized by fried egg with depressed center colonies, digitonin sensitive and negative to glucose fermentation and arginine utilization test. Out of 305 samples, 20 samples were selected for amplification by PCR technique using Mycoplasma 16S rRNA primer. Seven samples were positive to Mycoplasma species and gave amplified band at 1013 bp. subsequently, the seven isolates were sequencing. Four sequenced isolates (EGS1, EGL6, EGL1 and EGS2) were closely related to each other and very close to Mycoplasma bovis strains and far from M. leachii 99/014/6. Two sequenced isolates (EGL2 and EGL5) were closely related to each other and very close to Mycoplasma bovirhinis strains and far from M. leachii 99/014/6. One sequenced isolate (EGL3) was very close to Mycoplasma arginini strain and far from M. bovis 99/014/6. From these results, we conclude that conventional culture methods for diagnosis either by isolation and identification of Mycoplasma is a time-consuming method to diagnose mycoplasma infection So, these methods can be replaced by PCR and genome analysis technology.

Investigation of biotype, genotype and virulence associated genes inPasteurella multocida capsular type A strains from the respiratory tract of cattle and their relationship with disease cases

Pasteurella (P.) multocida, which causes respiratory disease in cattle, can also be found commensally in the upper respiratory tract. Therefore, it is epidemiologically important to determine the possible difference between pathogenic and commensal strains for the diagnosis of the disease. In this study, phenotypic and genotypic characteristics and virulence associated genes of P. multocida capsular type A strains isolated from the respiratory tract of sick and healthy cattle were investigated. Also, the relationship between the disease status of animals and the characteristics of the isolates were evaluated. Total of 51 P. multocida capsular type A strains were biotyped and genotyped by using commercial bacteria identification kits and PCR-fingerprint, respectively. Four major biotypes generated from xylose and sucrose test results were detected in 39 (76.47%) isolates. Eleven different biotypes were detected in the remaining 12 isolates (23.53%), which were separated from the 4 major biotypes by mannitol, indole, sorbitol, nitrate, VP, lactose, arabinose, salicin, and rhamnose test. Four genotypes were detected in P. multocida isolates and most of them (90.2%) had genotype I profile. Virulence associated genes, such as hgbA, exbB, exbD, tbpA, ptfA, hsf-2, tadD, nanB, nanH, pmHAS, sodA, sodC, and oma87 were detected in all isolates whereas hgbB, toxA, and hsf-1 genes were not found by PCR. PfhA gene were found in 49 isolates (96.07%) but, ompH gene could be detected only in 10 (18.86%) isolates. The findings of the study indicated that P. multocida capsular type A strains generated as xylose positive/sucrose negative (biotype II) and genotype II may be associated with respiratory diseases of cattle. However, ompH gene was observed to be associated with sucrose negative isolates, but not with disease cases. As a result, it was concluded that xylose positive/sucrose negative biotype and genotype II determined in P. multocida capsular type A strains isolated from respiratory diseases of cattle, might be important epidemiological criteria in the differentiation of commensal and pathogenic isolates.

Phenotypic and functional characterization of bovine mucosal associated invariant T (MAIT) cells

Abstract Mucosal associated invariant T (MAIT) cells are innate-like T cells that are prevalent in the blood and tissues of many mammalian species. MAIT cells recognize small metabolites derived from the riboflavin pathway that are presented by the highly conserved MHC-related protein-1 (MR1). MAIT cells have been described to play a proinflammatory role during several viral and bacterial diseases in humans and in rodent models. While MAIT cells have been described in cattle at the transcriptional level, little is known about the phenotype or function of this population in the bovine during steady-state or disease conditions. Here, using tetramers that bind to the MR1-restriced T cell receptors on MAIT cells, we investigated the frequency and phenotype of MAIT cells in the peripheral blood and bronchoalveolar lavage of healthy calves and animals with respiratory disease. We determined that cattle have 2 subsets of MAIT cells, defined by their expression of CD8; that bovine MAIT cells have the capacity to secrete IFNγ and IL-17 in response to mitogen stimulation; and that the frequency of MAIT cells in the bronchoalveolar lavage increases in the context of respiratory infection, suggesting that MAIT cells may play an important role in host defense in the respiratory tract of cattle.

Genotyping and antimicrobial resistance patterns of Mannheimia haemolytica, Pasteurella multocida and Histophilus somni isolated from the upper and lower respiratory tract of feedlot cattle

In the last 10 years, the prevalence of Pasteurellaceae resistant against drugs typically used for bronchopneumonia (BP) control has increased in feedlot cattle. Surprisingly, it is not clear whether this increase in the prevalence of multidrug resistant (MDR) bacteria is due to the spread of 1 or few multiple MDR clones among cattle during the feeding period (i.e. horizontal spread) or due to the recrudescence of MDR clones already present in the respiratory tract of cattle upon arrival at feedlots. Recently, we reported a high prevalence of MDR M. haemolytica, P. multocida, and H. somni isolated from cattle with BP in 4 feedlots in Western Canada. Unfortunately, as we did not genotype these isolates, it was not possible to determine whether a few or a large number of MDR clones were present in these feedlots, supporting either a horizontal spread of MDR clones among cattle or a recrudescence from carriers. Therefore, the objective was to genotype M. haemolytica, P. multocida, and H. somni isolates using pulsed field gel electrophoresis (PFGE).

Regulation of tracheal antimicrobial peptide gene expression in airway epithelial cells of cattle.

β-defensins are an important element of the mucosal innate immune response against bacterial pathogens. Tracheal antimicrobial peptide (TAP) has microbicidal activity against the bacteria that cause bovine respiratory disease, and its expression in tracheal epithelial cells is upregulated by bacterial products including lipopolysaccharide (LPS, a TLR4 agonist), Pam3CSK4 (an agonist of Toll-like receptor 2/1), and interleukin (IL)-17A. The objectives of this study were to identify the signalling pathway by which LPS, Pam3CSK4 and IL-17A induce TAP gene expression, and to determine the effect of glucocorticoid as a model of stress on this epithelial innate immune response. In primary cultures of bovine tracheal epithelial cells (bTEC), LPS, Pam3CSK4 and IL-17A each stimulated TAP gene expression. This effect was abrogated by caffeic acid phenylester (CAPE), an inhibitor of NF-κB. Similarly, western analysis showed that LPS, Pam3CSK4 and IL-17A each induced translocation of NF-κB p65 from the cytoplasm to the nucleus, but pre-treatment with CAPE inhibited this response. Finally, pre-treatment of bTEC with the glucocorticoid dexamethasone abolished the stimulatory effect of LPS, Pam3CSK4 and IL-17A on upregulation of TAP gene expression. These findings indicate that NF-κB activation is necessary for induction of TAP gene expression by LPS (a TLR4 agonist), Pam3CSK4 (a TLR2/1 agonist), or IL-17A. Furthermore, this stimulatory response is inhibited by glucocorticoid, suggesting this as one mechanism by which stress increases the risk of bacterial pneumonia. These findings have implications for understanding the pathogenesis of stress-associated bacterial pneumonia, and for developing methods to stimulate innate immune responses in the respiratory tract of cattle.

Comparative investigations of immune response of calves at different intervals between primary and secondary immunization using inactivated bovine herpes virus 1 vaccine

Bovine herpesvirus 1 (BHV-1) is one of the most siginificant causes of infections of the respiratory tract of cattle and immunoprophylaxis has a key role in curbing this infection. The intensity of the immune response against BHV-1 following immunization using inactivated commercial vaccines varies depending on the type of vaccine, but it is generally believed that they provide good protection from the development of the clinical form of the infection, and that they are safe. The paper present the development of the humoral immune response in fattening calves that were immunized against bovine herpesvirus 1 (BHV-1) at different time intervals between the primary and the secondary immunization. Calves were administered a commercial vaccine, and then they were divided into two groups which were revaccinated on days 14 or 21. Over a course of the 120 days of the duration of the experiment, blood and nasal mucus were sampled 11 times. The blood serum samples were examined for antibodies to BHV-1 using the virus neutralization (VN) test, and the nasal mucus samples were analyzed using the VN test and the ELISA method. Following revaccination, it was established that there was an increase in the antibody titer in blood of all experimental animals, and it was maintained at a high level up until the very end of the experiment (day 120). In the blood serums, maximum mean values for the antibody titer were determined on day 30 in the group that was revaccinated on day 14, and on day 45 in the group of calves revaccinated on day 21. In nasal mucus, antibodies were established at the earliest, using the virus neutralization test, on day 14 following vaccination, and using the ELISA method only after revaccination. The highest antibody titer in nasal mucus was established on day 45 in the group revaccinated on day 21, and on day 120 in the group revaccinated on day 14. Based on the established antibody titer values, calves can be revaccinated using the inactivated BHV-1 vaccine already on day 14.

Mycoplasmas isolated from the respiratory tract of cattle in Bosnia and Herzegovina

This study represents Mycoplasma species isolated from the respiratory tract of cattle in Bosnia and Herzegovina between 2002 and 2010. A total of 328 nasal swabs and 59 lung samples were submitted for isolation of mycoplasmas. Mycoplasmas were isolated from 27 samples (6.9%). M. bovis was recovered from eight nasal swabs and two lungs, while M. bovirhinis (n=4) and Acholeplasma sp. (n=1) were detected only in nasal swabs. Twelve mycoplasma isolates were unidentified (44.4%).

Bovine coronaviruses from the respiratory tract

Bovine coronaviruses (BoCV) have been associated with bovine respiratory diseases (BRD) in cattle pulled for treatment in feedlots and from healthy cattle in the US with BoCV identified by virus isolation from nasal swab and bronchoalveolar lavage (BAL) samples, and also by seroconversions for the detection of active infections. Bovine coronaviruses have been identified in pneumonic lungs from field cases, often in combination with other viruses and bacteria including Mycoplasma spp. Experimentally, BoCV infections have caused respiratory tract lesions. It has been suggested there is a dual tropism of BoCV for the respiratory and enteric tracts. Historically, BoCV was associated with neonatal enteric disease and adult winter dysentery. Respiratory isolates were identified later and focused on post-weaning cattle entering feedlots. Control measures are limited. There are USDA-licensed killed and modified live virus (MLV) vaccines for BoCV; however, these are for control of the enteric disease in cattle. There are no USDA-licensed vaccines for control of BoCV associated BRD in cattle. The purpose of the study was to compare BoCVisolates from the respiratory tract of cattle in Oklahoma to reference respiratory tract and enteric strains via antigenic and genetic procedures.

Mycoplasmas Isolated from the Respiratory Tract of Cattle and Goats in Tanzania

A microbiological study of the mycoplasma flora in the respiratory tracts of cattle and goats in selected regions of Tanzania is described. In the examination of cattle, mycoplasmas were isolated from 60 (17.8%) of the 338 examined lung samples, 8 (47.1%) of the 17 lymph nodes, 4 (13.3%) of the 30 pleural fluid samples and 4 (3.9%) of the 103 nasal swabs examined. All the isolates were identified as Mycoplasma mycoides subsp. mycoides, Small Colony type except for one isolate from pleural fluid which was identified as Mycoplasma arginini. M. mycoides subsp. mycoides, Small Colony type was isolated from samples originating from Dodoma, Iringa, Mbeya, Morogoro and Shinyanga regions where outbreaks of contagious bovine pleuropneumonia had been reported. In the examination of goats, mycoplasmas were isolated from 54 (34.0%) of the 159 examined lung samples, 41 (18.1%) of the 226 nasal swabs and 4 (40.0%) of the 10 pleural fluid samples. The species demonstrated were Mycoplasma capricolum subsp. capripneumoniae, M. mycoides subsp. mycoides, Small Colony type Mycoplasma ovipneumoniae and M. Capricolum subsp. arginini. The isolation of M. capripneumoniae in the Coast and Morogoro regions confirmed the presence of contagious caprine pleuropneumonia in the regions.

Induction of pulmonary immunity in cattle by oral administration of ovalbumin in alginate microspheres

Respiratory infectious diseases are an important cause of economic losses to the cattle industry. There is a need for an effective, easy to administer vaccine to the critical bacterial pathogens that cause pneumonia in cattle. An orally administered vaccine could be given to a large number of animals without significant stress to the animals and with minimal labor. The purpose of this study was to determine whether the oral administration of a model antigen (ovalbumin) in alginate microspheres could induce pulmonary immunity in cattle. Calves were vaccinated orally with ovalbumin (OVA) following either a subcutaneous (SC) or oral priming dose of OVA. Calves primed and boostered by oral administration (oral/oral) of OVA encapsulated in alginate microparticles had increased numbers of antigen-specific IgA ASCs (ASCs) in bronchoalveolar lavage (BAL) fluids. Calves that received a SC priming followed by an oral booster inoculation (SC/oral) of OVA in alginate microspheres had a greater number of anti-OVA IgA, IgG 1 and IgG 2 ASCs in BALF. SC/oral calves also had increased numbers of anti-OVA IgG 1 ASCs in peripheral blood whereas oral/oral calves had none. SC/oral calves had increased anti-OVA IgG 1, IgG 2, and IgA titers in BALF, and IgG 1 and IgG 2 in serum compared to both oral/oral and sham vaccinated calves. These results indicate that oral administration of antigen encapsulated in alginate microspheres results in a mucosal immune response in the respiratory tract of cattle. Furthermore, SC priming both enhanced the IgA response and stimulated an IgG 1 and IgG 2 response not seen in oral/oral calves. The difference in antibody isotype results suggest that design of the vaccination protocol can direct antibody responses as needed for a specific immunization program.

Globule leukocytes and bovine parasitic bronchitis.

Globule leucocytes (GL) occur in the respiratory tract of cattle from foetuses to adult animals. Large numbers of this cell have been found in the lungs of cows both in normal lung and in the lungs of cows from various outbreaks of respiratory diseases, but the significance was not known. In the rat and in cattle, the occurrence of GL in the gastrointestinal and urinary tracts has been associated with parasitic infections. The relationship between the respiratory tract GL and Dictyocaulus viviparus infections in cattle was studied in three groups of parasite free calves experimentally infected with D. viviparus. Group 1 animals received single challenge infections. Group 2 had three subgroups (2a, 2b, 2c) which were either vaccinated and challenged (2a), given a single challenge (2b) or used as vaccinated controls (2c). Group 3 with four subgroups (3a, 3b, 3c, 3d) were either used as environmental controls (3a), vaccinated and repeatedly challenged (3b), given normal larvae of D. viviparus and repeatedly challenged (3c) or as a trickled group 3d. Although GL were found in infected calves, it was not possible to demonstrate a clear cut correlation between their presence or density and the method of exposure.

An in vivo study of a glycoprotein gIII-negative bovine herpesvirus 1 (BHV-1) mutant expressing β-galactosidase: Evaluation of the role of gIII in virus infectivity and its use as a vector for mucosal immunization

We constructed a recombinant BHV-1 in which the glycoprotein gIII gene was replaced by the Escherichia coli lacZ gene. The resultant virus mimics the simple gIII deletion mutant in its growth characteristics in cell culture; however, it expresses β-galactosidase in virus-infected cells. Further haracterization of its virulence and the immune responses elicited by it was conducted in cattle. The mutant virus retained the ability to establish an infection when administered intranasally. Infected animals were also capable of transmitting virus to sentinel penmates. However, the mutant virus showed a reduced replication efficiency in the respiratory tract of cattle, as manifested by significantly lower virus shedding and a shorter duration of shedding when compared to wild-type (wt) BHV-1 infections. The mutant virus induced an efficient anti-BHV-1 antibody response and convalescent cattle were fully protected from subsequent wt virus challenge. In addition, cattle infected with the lacZ-expressing virus developed antibodies to β-galactosidase. Our results demonstrate that the presence of gIII is not a prerequisite for BHV-1 infection; however, gIII does play an important role in maintaining virus replication efficacy in its natural host. With respect to developing BHV-1 as a vaccine vector, our results indicate that deletion of the gill gene, which partially attenuates the virus and serves as a vaccine virus marker, does not compromise immunogenicity to BHV-1. Most importantly, this vector is effective in delivering foreign antigens to mucosal surfaces of the respiratory tract.