Calf Kidney Research Articles

CHARACTERISTICS OF THE EQUINE RHINOPNEUMONIA VIRUS AK-2011

Equine rhinopneumonitis is an acute, highly contagious disease found virtually worldwide. The object of the studies was the AK-2011 strain isolated from the horses suffering from rhinopneumonitis during an outbreak of abortions. The viability of the AK-2011 strain was assessed using a continuous line of calf trachea cells, a continuous line of calf kidney cells, a continuous line of sheep kidney cells, a continuous line of bovine kidney cells, a continuous line of green monkey kidney cells, a continuous line of Syrian hamster kidney cells, a primary trypsinized culture of horse kidney cells grown in tubes and flasks, and the AK-2011 laboratory strain of equine rhinopneumonitis virus with biological activity of 6.0 lg TCID50/cm3. Sequencing and polymerase chain reaction (PCR) analysis were performed. The virus isolated from the ORF68 gene in Kazakhstan appeared to be the most similar to the T-953 and 2222-03 strains isolated in the USA and Australia, respectively, in terms of phylogenetics. As to primary infections, cytopathic effects (CPE) induced by the AK-2011 virus stain (dilution 101) in calf trachea and horse kidney cell cultures were stable from the 1st to 10th passages, with biological activity of 5.75-6.00 lg TCID50/cm3. CPE caused by the virus were apparent on days 2-3, further developed intensively, and extended to 60-80% of the cell monolayer on days 5-7. In calf kidney, sheep kidney, green monkey kidney, and bovine kidney cell cultures, the same changes were observed 1-2 days later. The changes were slow, and by day 7-10 CPE extended to no more than 30-50% of ​​the cell monolayer. An attenuated strain AK-2011 of equine rhinopneumonitis virus was obtained. It was considered a candidate for the manufacture of a vaccine for equine rhinopneumonitis. Thus, the attenuated strain AK-2011 of the equine rhinopneumonitis virus was characterized.

ISOLATION AND IDENTIFICATION OF ROTAVIRUS FROM NEONATAL CALVES

Bovine rotavirus was successfully isolated in vero cell line and secondary calf kidney cell culture from faecal samples of newly bom calves affected with diarrhoea. The isolated viruses were cultivated from cell culture produced cytopathogenic effect which consisted of cytoplasmic granulation degeneration and detachment of infected cells from monolayer cell culture. The rotavirus was identified by indirect immunofluorecent test (IFT) and neutralization test (NT) by using reference hyperimmunser.

298 Late Gestational Nutrient Restriction of Primiparous Beef Heifers Decreases Neonatal Calf Body Weight and Organ Mass

Abstract We hypothesized that late gestational nutrient restriction (NR) would decrease organ mass in neonatal calves. Fall-calving crossbred beef heifers (BW: 466 ± 30 [SD] kg; BCS: 5.4 ± 0.5) bred to a single sire were individually fed 100% (control; n = 10) or 70% (nutrient restricted; NR; n = 12) of NASEM metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from d 160 of gestation to calving. Diets were based on chopped sorghum sudan hay and supplemented to meet targeted nutritional planes. Calves were removed immediately post-calving and fed colostrum and milk replacer relative to BW until euthanasia (48.7 ± 0.4 h of age) for organ mass determination. Nutritional plane was a fixed effect, and maternal sire, calf sex (when P < 0.25), and date of study initiation were covariates. Post-calving, NR dams were 62 kg BW and 2.1 BCS less (P < 0.001) than controls. Calf BW, liver, large intestine, omental/mesenteric fat, kidney, and heart weights were less (P ≤ 0.05) and carcass, rumen/reticulum/omasum, stomach complex, and total gastrointestinal tract weights tended to be less (P ≤ 0.09) in offspring from NR dams. Maternal nutrient restriction did not affect (P ≥ 0.11) weights of the small intestine, pancreas, abomasum, spleen, perirenal fat, adrenals, lungs, femur, and brain. Expressed per BW, lung weight was greater (P = 0.05), and brain and femur weight tended to be greater (P ≤ 0.10) in calves from NR dams, but other organ masses were unaffected (P ≥ 0.15). Expressed relative to brain weight, liver and large intestine weights were less (P ≤ 0.04) and carcass, total gastrointestinal tract, omental/mesenteric fat, and kidney weights tended to be less (P ≤ 0.08) in NR. In summary, late gestational NR decreased some organ masses in neonatal calves while sparing other tissues via asymmetric growth.

AK-2011 strain for the development of a vaccine against equine rhinopneumonitis.

Equine rhinopneumonitis is an acute, highly contagious disease found virtually worldwide. The purpose of the studies presented in this paper is to develop a technology for the manufacture of a cell-derived equine rhinopneumonitis vaccine, as well as to assess the safety and immunogenicity of the newly developed vaccine in laboratory animals model. The object of the studies was the AK-2011 strain isolated from the horses suffering from rhinopneumonitis during an outbreak of abortions. The viability of the AK-2011 strain was assessed using a continuous line of calf trachea cells, a continuous line of calf kidney cells, a continuous line of sheep kidney cells, a continuous line of bovine kidney cells, a continuous line of green monkey kidney cells, a continuous line of Syrian hamster kidney cells, a primary trypsinized culture of horse kidney cells grown in tubes and flasks and the AK-2011 laboratory strain of equine rhinopneumonitis virus with biological activity of 6.0lg TCID50/cm 3 . Sequencing and polymerase chain reaction analysis were performed. The virus isolated from the ORF68 gene in Kazakhstan appeared to be the most similar to the T-953 and 2222-03 strains isolated in the USA and Australia, respectively, in terms of phylogenetics. As to primary infections, cytopathic effects (CPEs) induced by the AK-2011 virus stain (dilution 101 ) in calf trachea and horse kidney cell cultures were stable from the first to tenth passages, with biological activity of 5.75-6.00 lg TCID50/cm 3 . CPEs caused by the virus were apparent on days 2-3, further developed intensively and extended to 60-80% of the cell monolayer on days 5-7. The vaccine results can be used to immunize horses on farms against rhinopneumonia, and horses should be immunized twice with an interval of 2-3 months.

Study of isolate of Infectious rynotracheitis virus identified in the acute course of the disease

Isolation of the virus from biological material from a two-month-old calf with pathology of the respiratory system from a herd with a morbidity rate of 48% was performed. After detection the presence of IRT antigens in the lungs of the dead animal, the pathogen was isolated on a continuous culture of calf kidney cells, where a characteristic cytopathic effect was observed. The genetic material of the bovine herpesvirus type 1 (Bovine herpesvirus-1, BHV-1) was identified by polymerase chain reaction in the test sample. The virus isolate was adapted to continuous cell cultures of calf kidneys, sheep kidney, cow embryo lung and calf trachea, and the most suitable biological system was determined, where adsorption and reproductive properties of the virus were more pronounced. It was found that the highest titer of infectious activity of BHV-1 isolate (6.1 lg TCD50/cm3) was obtained on continuous culture of lung embryonic cells of a cow embryo after its reproduction during three consecutive passages (observation period)

Design and study of immunogenicity of virus vaccine against viral pneumoenteritis of calves

The immunogenicity of a virus vaccine against viral pneumoenteritis in young cattle was studied in laboratory animals. The results of the selection of optimal vaccine strains to study the effects of different inactivants on infectious rhinotracheitis virus, viral diarrhea, parainfluenza-3, respiratory syncytial virus, rotavirus and coronavirus are presented. The accumulation of avirulent vaccine virus strains was carried out using established virological methods on transplanted cell cultures of MDBC (calf kidney cells) and SPEV (fetal pig kidney cells). Theotropine and formalin were used as inactivating agents to develop inactivation regimes for vaccine strains - components of the experimental vaccine. The antigenic activity of attenuated strains of infectious rhinotracheitis virus, viral diarrhea, parainfluenza-3, respiratory syncytial virus, rotavirus and coronavirus on white mice and calves and the level of specific antibodies in the sera of guinea pigs immunized with vaccines using different adjuvants were studied. Avirulent strains of viruses were used in the design of a new virus vaccine against viral pneumoenteritis: infectious rhinotracheitis (IBR-VBF-VHAVM No. 404); diarrhea (VDVBF-VHAVM No. 406); parainfluenza-3 (PG-VBF-VHAVM No. 403); respiratory syncytial virus (RSV-VBF-VHAVM No. 405); rotavirus (RTV-VBF-VHAVM No. 401) and coronavirus (CV-VBFVHAVM No. 407). The selected vaccine strains are non-reactive and induce active production of antiviral antibodies in sufficiently high titres in both laboratory animals (white mice) and farm animals (cattle). The most effective inactivating agents are 0.1% theotropine and 0.2% formalin. Two types of oil-based adjuvants, ISA 15 and ISA 25, were used to select the optimal adjuvants for the design of the virus vaccine. Adjuvant ISA 15 was used at 15% of the antigen quantity, ISA 25 at 25%. The adjuvant IZA 15 at a concentration of 15% is the optimum adjuvant for the preparation of an experimental virus vaccine against viral pneumoenteritis in young cattle.

COMPARATIVE STUDY BETWEEN ROTA VIRUSES ISOLATED FROM DIARRHEATIC INFANT BABIES AND NEWLY BORN CALVES

Rota viruses were isolated in vero continuous cell line from infant babies and newly borne calves affected by diarrhea after treatment in every passage with 10 mg/ml of trypsin and adding 0.5 mg/ml of trypsin in the maintenance media. The isolated viruses induced distinctive and progressive type of cytopatheic effect in infected cells and highest titer was 2x10 6. TCID,50/0.1 for bovine viral isolate. The isolated 50/0.1 for human viral isolate and 2x10 » viruses were identified by indirect fluorescent antibody technique by using reference calf rotavirus antisera. Comparative study were conducted on both human and bovine viral isolate including growth in different cell culture, vero cell line was very sensitive to support growth of both viruses than secondary embryonic calf kidney cell culture, lamb testes and primary embryonic chicken fibroblast cell culture. Both viruses induced morphologically similar kind or plaques in vero cell line but plaqes formed by human isolate were larger in size about 1.5 - 2.5 mm in diameter than those of bovine isolate 1.5 - 2 mm. Cross reactive viral antigens were detected between the isolated viruses in indirect fluorescent antibody technique but not in serum neutralization test by using reference bovine rotavirus antisera.

Ізоляція коронавірусу ВРХ в культурах клітин

One of the most common viruses in the world that causes disease in cattle is the bovine coronavirus (BCoV). This virus is the causative agent of respiratory and gastrointestinal infections in newborn calves, resulting in significant economic losses in both dairy and meat farming. Considering the complex epizootic situation with the coronaviruses in the world and partial antigenic affinity of BCoV with coronaviruses of other species of animals and humans, the isolation of new strains of coronaviruses, their identification and optimization of cultivation conditions becomes extremely important and relevant. The aim of our research was to determine the features of methods of isolation of bovine coronavirus and to select methods for its cultivation in cell culture in order to obtain the virus with the highest titers of infectious activity. Isolation of BCoV was performed in monolayers of MDBK and the primary-trypsinized calf kidney culture cells, using 20 samples collected from calves with clinical signs of respiratory or/and gastrointestinal disease. 16 samples were positive for BCoV by means of Real-Time PCR test. Up to fifth serial passage, only 4 of these isolates presented typical syncytial cytopathic effect. It has been experimentally established that the continious calf kidney cell culture line (MDBK) and the primary-trypsinized calf kidney culture (CK) are suitable for BCoV isolation and accumulation. The infectious titer of bovine coronavirus at the level of the fifth passage in the cultures of MDBK and CK cells reached 5.54 ± 0.16 lg TCD50/ml and 5.59 ± 0.14 lg TCD50/ml, respectively. However, due to the high cost of obtaining primary-trypsinized cell cultures, this isolation method may be unacceptable to most pharmaceutical companies and laboratories. Also after 5 serial passages, the viral material was again examined in Real-Time PCR to confirm the isolation of BCoV - the study of 4 samples with a characteristic syncytial CPE had a positive result in Real-Time PCR. However, of the Real-Time PCR-positive 12 samples, the virus could not be isolated in continuous cell cultures of MDBK and Vero, as well as in primary-trypsinized cattle lung and kidney cell cultures. This fact may indicate the presence of different strains of BCoV circulation in farms in our country. Further research is planned to be focused on optimizing the methods and modes of BCoV strains isolation, as well as to identify and study the cultural properties of new strains of BCoV circulating in Ukraine. We will also continue the study of the obtained viral isolate for the subsequent development of tools for the diagnosis and immunoprophylaxis of coronavirus infection in veterinary medicine.

Isolation of Bovine leukemia virus from cows with persistent lymphocytosis in Iraq

This is the first study to report on the isolation of bovine leukemia virus (BLV) from peripheral blood mononuclear cells of two cross bred cows in Iraq. The cattle were seropositive by ELISA when selected while being surveyed for the detection of BLV. Among six cows, two were cases of persistent lymphocytosis (PL). Cytopathology was characterized by the formation of multinucleated giant cells (syncytia) and cytoplasmic vacuoles. Moreover, the viruses produced clear plaques on the monolayer of the primary fetal calf kidney (FCK) cells. Inhibition of plaque formation by BLV-antisera suggested a diagnosis of BLV, which was further confirmed by PCR. Cells infected with the isolates were positive to a monoclonal antibody against the viral gp51 trans-membrane glycoprotein by immunocytochemistry. Both isolates replicated and induced cytopathic effects in bovine and human cell line cultures. Phylogenetic analysis based on partial gp51 env gene sequences revealed that Iraqi strain highly homogenous with Turkey strain (100%) and had 1% distance value with other world strains. In conclusion, this present study found that BLV-infected cattle with PL can be a source for viral isolation, and the cytopathological features of the virus infection are arranged and differ depending on the cell type. This is the first study to report on the isolation of the EBL virus in Iraq, and it provides the basis for further studies about a BLV Iraqi strain that can help control this disease.

Визначення цитотоксичності антимікробних препаратів і дезінфектантів на культурі клітин нирки теляти

Goal. To study the cytotoxicity of certain antibacterial preparations used in the treatment of infectious diseases of the urinary system, as well as some disinfectants for calf kidney cell culture. Methods. Cytological, toxic effects test on primary calf kidney cell culture. Determination of the maximum permissible concentrations of antibiotics, antiseptics, and disinfectants that do not yet lead to degeneration of the cell monolayer of calf kidney cells. Results. Tests of some drugs that are often used in veterinary medicine show that the least toxic to calf kidney cells of all tested antibacterial agents are Oxymicol 20 (active substance — oxytetracycline base), Norsulfazole sodium, and Farmazin 50 (active substance — tylosin). Conclusions. These drugs or similar with the same active substances should be used in the treatment of infectious diseases of the kidney and urinary system of cattle and other animals. The possibility of using the method of cultivation and use as test objects of primary cultures of eukaryotic cells in the selection of the optimal composition of therapeutic and prophylactic drugs or disinfectants for veterinary medicine. The obtained results of cytotoxicity can help veterinarians in choosing antimicrobials and disinfectants in terms of their wide range on the market.

Optimization of cultivation parameters for bovine respiratory syncytial virus strain Vologda/2019

There are currently many controversial issues in the study of bovine respiratory syncytial infection. In this regard, it is relevant to study the biological properties of the virus, optimize the methods of its cultivation and select the most technologically advanced methods of designing diagnostic and prevention tools for this disease. The aim of this work was to select sensitive cell systems and to optimize the cultivation parameters in selected cell cultures. The Vologda/2019 strain of the bovine respiratory syncytial infection virus isolated from biological material obtained from a calf with respiratory symptoms was used in the experiment. The strain was adapted to the continuous cell culture derived from bovine turbinate tissue (BT) and deposited in the State collection of microorganism strains at FGBI “ARRIAH”. It was established that the continuous cell lines of fetal bovine trachea (FBT) and calf kidney (RBT) are the most sensitive cell systems for the reproduction of the bovine respiratory syncytial virus strain Vologda/2019, the maximum accumulation of the virus was observed in these cell cultures. The cytopathic activity of the virus in the FBT cell culture ranged from 4.78 ± 0.18 to 5.50 ± 0.16 lg TCID 50 /cm 3 , and in the RBT cell culture – from 4.00 ± 0.23 to 4.75 ± 0.20 lg TCID 50 /cm 3 at days 4–5 of cultivation. It was determined that in case of multiplicity of inoculation of FBT and RBT cell cultures with the virus at 0.1 lg TCD 50 /cell and the use of 2% glutamine in the maintenance nutrient medium, as well as 2% horse or cattle blood serum, it is possible to obtain virus material with high cytopathic activity.

Genetic characterization of Shigatoxigenic and enteropathogenic Escherichia coli O80:H2 from diarrhoeic and septicaemic calves and relatedness to human Shigatoxigenic E. coli O80:H2.

The purpose of this work was to identify and genetically characterize enterohemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) O80:H2 from diarrhoeic and septicaemic calves in Belgium and to comparing them with human EHEC after whole genome sequencing. Ten EHEC and 21 EPEC O80 identified by PCR between 2009 and 2018 from faeces, intestinal content and a kidney of diarrhoeic or septicaemic calves were genome sequenced and compared to 19 human EHEC identified between 2008 and 2019. They all belonged to the O80:H2 serotype and ST301, harboured the eaeξ gene, and 23 of the 29 EHEC contained the stx2d gene. Phylogenetically, they were distributed in two major sub-lineages: one comprised a majority of bovine EPEC whereas the second one comprised a majority of stx2d bovine and human EHEC. Not only EPEC but also EHEC O80:H2 are present in diarrhoeic and septicaemic calves in Belgium and are genetically related to human EHEC. These findings support the need to assess cattle as potential source of contamination of humans by EHEC O80:H2 and to understand the evolution of bovine and human EHEC and EPEC O80:H2.

CULTURAL PROPERTIES OF BOVINE RESPIRATORY SYNCYTIAL VIRUS STRAIN AB1908

Cattle respiratory diseases are some of the most spread pathologies that can cause economic damage, resulting from fi nancial losses and costs of treatment and diagnostics. One of the major factors contributing to respiratory pathology development is bovine respiratory syncytial infection. The analysis of serological testing, performed by the FGBI “ARRIAH” Reference Laboratory for Cattle Diseases in 2017–2018, showed that respiratory syncytial virus seroprevalence in animals of dairy farms is 60%. Herewith, it was noted that the most susceptible animals to this infection are calves under one year of age. The eff ectiveness of bovine respiratory syncytial infection control measures depends on timely diagnosis; that is why reliable and accurate diagnostic tools are needed, including optimal techniques of virus isolation from pathological material. For successful virus isolation from clinical samples, it is necessary to adhere strictly to optimal parameters of this agent cultivation. This paper presents data on study of bovine respiratory syncytial virus strain AB 1908 cultural properties. The tests performed showed that a continuous bovine turbinate (BT) cell line, continuous bovine fetal trachea (FBT) cell line and continuous bovine calf kidney (RBT) cell line are sensitive for cultivation of this agent and can be used to prepare viral suspension, needed for further research. Virus titre in BT cell culture was 4.33 ± 0.16 – 4.66 ± 0.12 lg TCID50/ cm3, in RBT cell culture – 4.33 ± 0.33 – 4.7 ± 0.36 lg TCID50/cm3 and in FBT cell culture – 4.13 ± 0.20 – 4.78 ± 0.17 lg TCID50/cm3. The following virus cultivation optimal parameters were also determined during this study: the age of the culture for virus inoculation should be 1–2 days and multiplicity of inoculation should be 0.1 TCID50/cell.

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A loop-mediated isothermal amplification assay for rapid and sensitive detection of bovine papular stomatitis virus

Bovine papular stomatitis virus (BPSV) causes pustular cutaneous disease in cattle worldwide. This paper describes the development of a specific loop-mediated isothermal amplification (LAMP) assay to detect BPSV which did not cross-react with other parapoxviruses. To assess analytical sensitivity of this LAMP assay, DNA was extracted from serially diluted BPSV from which the infectious titer was determined by a novel assay based on calf kidney epithelial cells. The LAMP assay had equivalent analytical sensitivity to quantitative PCR, and could detect as few as 86 copies of viral DNA per reaction. These results suggest that the assay is a specific and sensitive technique to rapidly diagnose bovine papular stomatitis in domestic animals.

Electron Microscopy Fixative Fails to Inactivate Rift Valley Fever Virus

Rift Valley fever (RVF) virus, a member of family Bunyaviridae, genus Phlebovirus, is a mosquito‐borne zoonotic pathogen endemic to Sub‐Saharan Africa and the Arabian Peninsula that causes severe epizootics in ruminants. RVF outbreaks in ruminants are associated with mass abortions and high mortality in younger animals. Human infections at all ages are associated with acute febrile illness that may progress to more severe disease including encephalitis and severe hepatitis leading to fatal hemorrhagic fever. RVF virus (RVFV) is classified as an overlap select agent by the Center for Disease Control and Prevention and the US Department of Agriculture. In the US, work with virulent RVFV strains must be conducted in BSL‐3+ and requires RVFV specific select agent clearance.In conjunction with ruminant challenge model development for evaluation of new RVFV vaccines and therapies, we have been examining the pathology of RVFV in multiple organs. Our initial histopathology findings warranted examination of infected tissues by transmission electron microscopy (EM). Consequently, during early time‐point post‐infection calf necropsies, we collected 2–5 mm cube sized samples of calf kidney, liver and spleen in room temperature (RT) 2.5% glutaraldehyde in Sorenson's buffer (glut), a standard EM fixative that preserves morphology for EM. Glutaraldehyde shares fixation characteristics with 10% neutral buffered formalin that is already approved for use in our BSL‐3Ag facility. Post ~2 hours at RT, samples were stored at 4° C, as per advice from our EM experts, for a minimum of 7 days. Because glut was a new fixative for our facility, we conducted proof of inactivation tests on our samples. RVFV PCR‐positive, unfixed tissue from each organ of interest from one of the calves and PBS were used as positive and negative controls respectively. Samples were rinsed in PBS, homogenized and blind passaged 3 times using standard virus propagation protocols. The cell monolayers were checked post passage for cytopathic effect (CPE) and standard plaque assays were conducted after the third passage. Additionally, in a separate experiment, using full strength and serially diluted glut in a no virus simulated viral propagation protocol, we tested for a direct toxicity effect of glut on our Vero cells during the initial 1 hr incubation time‐period.Plaque formation was observed on cells incubated with glut samples containing RVFV‐positive tissues and positive control tissues but not on cells incubated with PBS or glut. This indicates that the glut treated tissue samples still contained viable RVFV. Currently, we have experiments planned to consider the role of temperature in glut viral inactivation.Support or Funding InformationThis work was funded by grants of the Department of Homeland Security Center of Excellence for Emerging and Zoonotic Animal Diseases (CEEZAD), Grant No. 2010‐ST061‐AG0001, USDA Agricultural Services project 3020‐32000‐005‐00D and the Kansas Bioscience Authority.

Peptide Regulation of Cells Renewal Processes in Kidney Tissue Cultures from Young and Old Animals.

Polypeptide complex isolated from calf kidneys stimulates the processes of cell renewal in organotypic kidney tissue cultures from young and old rats. The polypeptide complex enhances expression of proliferation marker Ki-67 and reduces expression of proapoptotic peptide p53 in kidney explants obtained from young and old animals. Short peptides T-31 (AED) and T-35 (EDL) also stimulate proliferation and reduce apoptosis of the kidney cells, but to a lesser degree than the polypeptide complex. The results provide the basis for further investigation of the polypeptide complex as a preparation for the therapy of kidney diseases, including age-related pathologies.

Features of the replication of hantaviruses - pathogens of hemorrhagic fever with renal syndrome in cell cultures of different origin

There are presented results of the adaptation ofpathogens of hemorrhagic fever with renal syndrome (HFRS) virus Puumala and Dobrava strains, to the reproduction in cell cultures of the simian origin - Green monkey kidney (Vero, 4647), Green monkey spleen (455), calf kidney (PT-1) and sheep kidney (4184). Hantaviruses were shown to be able to adapt to the replication in the new cellular system differing in species origin from the culture of Vero E6 origin in which the strains were isolated. This fact demonstrates the heterogeneity of hantavirus population, with the existence of «quasispecies» which can recognize and bind to ligands (b3 integrins) of the other species specificity. The highest yield of the virus, which is comparable with the culture of Vero E6, culture Vero provides. In Vero cells after 3-4 passages Puumala and Dobrava-Belgrade viruses replicate also actively as in a permissive culture Vero E6, that proposes the use of Vero culture cell allowed for the production of immunobiological drugs administered to people as a substrate in the delivery of vaccines against HFRS.

Peptides regulate expression of signaling molecules in kidney cell cultures during in vitro aging.

We studied the effect of polypeptide complex isolated from calf kidney and short peptides T-31 (AED) and T-35 (EDL) on the expression of signaling molecules, markers of cell renewal (Ki-67, p53), remodeling of the extracellular matrix (MMP-14), and immune response (IL-8) in primary kidney cell cultures during aging. The complex of renal polypeptides and T-31 peptide activate cell renewal processes during aging of the renal epithelium, while gelatinase MMP-14 is the target of T-35 peptide.

Short communication. First report in Cuba of bovine coronavirus detection in a winter dysentery outbreak

Bovine coronavirus (BCoV) infection causes epidemics of acute diarrhea in calves and winter dysentery (WD) in adult cattle. The disease in adult cattle can cause a decrease in milk production resulting in serious economic losses. In Cuba, BCoV infections have not been previously reported. During 2004, many outbreaks of enteric disease have occurred in adult cattle from thirteen dairy farms in the central and western part of the island. The clinical features of the outbreaks resembled those of WD, such as decrease in milk production and, 24 hours later, diarrhea, sometimes bloody, that lasted until the animals recovered in about 7-15 days. Laboratory confirmation of BCoV infection was provided by hemagglutination test (HAT) in 24 samples from four dairy farms from three provinces; RT-PCR assays confirmed the presence of BCoV in three of these samples. Cell culture isolation in secondary calf kidney cells was obtained from four pools of fecal diarrheic samples from each dairy farm. It was remarkable that the disease was also observed during summer. Studies of molecular characterization of the viral strain are in progress.