Malignant catarrhal fever (MCF) is a rare, under-explored lethal viral infection of cattle with gammaherpesvirus aetiological agents. Most often, the disease occurs on farms where cattle and sheep are kept together. However, other trigger mechanisms and environmental factors contribute. This study investigates the causation of MCF. An outbreak of MCF occurred in June - August 2017 in Kharchev village in Irkutsk Oblast, Russia. In this paper, we provide epidemiological (sanitary status of pastures, watering places, and premises) and weather data during the outbreak, and descriptions of the clinical signs and post-mortem changes in cattle. The virus was detected and isolated from pathological material samples and identified by molecular methods. Extreme weather conditions, mixed-herd cattle and sheep farming, and unsatisfactory feed quality contributed to the outbreak. A virus related to herpesvirus OvHV2 was isolated and typed (MCF/Irkutsk/2017). Phylogenetic analysis showed its close genetic relationship to isolates from cattle and sheep in Germany, USA, and the Netherlands. Sporadic outbreaks of MCF caused by biotic and abiotic factors together are typical for the Russian Federation, and the Irkutsk outbreak epitomised this. Temperature anomalies caused pasture depletion, resulting in feed and water deficiency for grazing animals and dehydration and acidosis. Heat stress in animals ultimately led to the occurrence of MCF in the herd.

The clinicopathologic manifestations of bovid herpesvirus-4 (BHV-4; FCAHV strain)-induced infection of the lower portion of the urinary tract were characterized in 12 adult neutered male and 6 female specific-pathogen-free cats, and were compared with those in 12 neutered male control cats. Six neutered male and 6 female cats were given immunosuppressive doses of methylprednisolone acetate prior to inoculation of their urinary bladders with BHV-4. Six neutered male control cats were given immunosuppressive doses of methylprednisolone acetate prior to inoculation of their urinary bladders with uninfected tissue culture control inoculum. Six additional neutered male control cats were exposed only to uninfected tissue culture control inoculum. All cats were observed for 90 days following inoculation. Dysuria and gross hematuria were observed in only 1 BHV-4-exposed cat. Radiographic abnormalities of the lower portion of the urinary tract were not observed. Microscopic hematuria, crystalluria, and lipiduria were identified with similar frequency in BHV-4-exposed and control cats. Results of urine culturing for bacteria, mycoplasma, ureaplasma, and viruses were negative. Viruses were not isolated from blood leukocytes collected from exposed or control cats. Three to 6 weeks after inoculation, high concentrations of BHV-serum 4 antibodies were detected in all exposed cats by an indirect fluorescent antibody test. Light microscopic examination of the urinary tract revealed multifocal lymphoid cystitis in 2 BHV-4-exposed cats. Except for suppurative bronchitis in 1 BHV-4-exposed cat given glucocorticoids, morphologic differences in urinary and extraurinary tissues were not observed. In urinary bladder tissue collected 90 days after inoculation, BHV-4 was reisolated from urinary bladder explants of all but 1 exposed cat. Virus was also isolated from a kidney explant of 1 exposed male cat, and spleen cell cocultures of 1 exposed female cat given glucocorticoids. Bovid herpesvirus-4 (FCAHV strain) caused persistent urinary tract infections in male and female specific-pathogen-free cats. Detection of occult BHV-4 infection required isolation of virus from tissues by explantation, or demonstration of specific BHV-4 antibodies by immunofluorescent fluorescent techniques. Administration of glucocorticoids prior to inoculation did not enhance morbidity associated with BHV-4 urinary tract infection. Further investigations are needed to determine the pathogenic role of BHV-4 in 4 noninduced feline lower urinary tract disease.

In vitro activities of 9-([2-hydroxyethoxy] methyl) guanine (acyclovir), (E)-5-(2-bromovinyl)-2'deoxyuridine, 9-beta-D-arabinofuranosyladenine (vidarabine), 5-iodo-2'-deoxyuridine (idoxuridine), and 5-trifluoromethyl-2'-deoxyuridine (trifluridine) were studied against 6 strains of feline herpesvirus-1. A significant difference was not detected among viral strains in their susceptibility to these compounds (P = 0.442). The relative potency of these compounds was trifluridine greater than greater than idoxuridine greater than virdarabine greater than bromovinyldeoxyuridine greater than greater than acyclovir. Concentrations of trifluridine and idoxuridine (0.67 and 6.8 micromole, respectively) required to reduce plaque numbers by 50%, compared with that of controls, were significantly lower (P less than 0.001) than were those of other compounds.

To obtain synchronous infection, 10 cats were inoculated with feline herpesvirus-1 (FHV-) on the oral, nasal and conjunctival mucosa. Swab specimens of the nasal conjunctival, and pharyngeal mucosa were obtained for virus isolation from each cat before inoculation and at 3-day intervals thereafter until postinoculation day 21. Recovery of virus and evidence of clinical signs were used to document FHV-1 infection. Serum was obtained from blood samples collected sequentially from each cat between day 0 and postinoculation day 90. Virus-neutralizing antibody titer was determined in all serum specimens. Immunoprecipitation with [35S]methionine- and [14C]glucosamine-labeled viral antigens, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was performed on each specimen. Three precipitation bands with approximate molecular weights of 105,000, 68,000, and 60,000 were separated from [14C]glucosamine- and [35S]methionine-labeled immunoprecipitates. The concurrent detection of virus-neutralizing antibody glycoprotein-specific immunoprecipitins implied that in cats, the FHV-1 glycoproteins were important in the induction of virus-neutralizing antibodies to FHV-1.

Cerebrospinal fluid (CSF) changes are significant for antemortem diagnoses of some neurological diseases. The aim of this study was to evaluate if the concentration of L-lactate in CSF could be used to differentiate healthy from encephalitic cattle. Cerebrospinal fluid samples from healthy cattle (n = 10) and from those naturally affected by rabies (n = 15), bovine herpesvirus type 5 meningoencephalitis (n = 16), histophilosis (n = 6), or bacterial encephalitis (n = 4), including 1 case of listeriosis, were collected and analyzed. Physical, biochemical (i.e., protein and glucose), and cellular analyses were performed in fresh samples. L-lactate, electrolytes (sodium, potassium, and chloride), calcium, and magnesium concentrations were measured in CSF samples that were kept frozen. L-lactate concentrations were also measured in plasma. Analysis of variance was used for comparison between groups and receiver operating characteristic analysis was performed considering L-lactate in CSF of healthy versus encephalitic cattle. The CSF L-lactate concentration was significantly higher in cattle with bacterial encephalitis than in healthy cattle; however, it did not differ between viral and bacterial encephalitis. The calcium concentrations were lower in cattle with encephalitis. L-lactate concentration in CSF > 3.6 mmol/L can be accepted as a cut-off value to indicate encephalitis. Thus, L-lactate in CSF is important for the diagnosis of encephalitis in cattle. Despite the small number of cases of bacterial encephalitis, it is suggested that L-lactate was not important for the differentiation between viral and bacterial encephalitis. Additional studies with a greater number of observations are necessary to clarify this, specifically in cases of listeriosis.

Clinicopathologic manifestations of induced infection of the feline lower urinary tract with bovid herpesvirus-4 (BHV-4, strain FCAHV) were characterized in 6 conventionally reared adult cats (2 sexually intact males, 2 castrated males, and 2 females). Two additional control cats were exposed with noninfected cell culture control inoculum. Clinical and radiographic signs of lower urinary tract disease were not observed in exposed or control cats. Microscopic hematuria was detected in urine samples collected by cystocentesis from 4 of 6 exposed cats and 1 of 2 control cats. Results of culture of urine for bacteria, mycoplasmas, ureaplasmas, and viruses were consistently negative. Low titer of serum BHV-4 (strain FCAHV)-neutralizing antibodies was detected in 4 of 6 exposed cats, but not in controls. Gross abnormalities of the urinary tract were not observed in any cat. Light microscopic examination of serial sections of the lower urinary tract revealed mild focal lymphoid cystitis in 2 of 6 exposed cats, one of which also had increased amounts of connective tissue and proliferation of blood vessels in the urinary bladder lamina propria. Ninety days after initial exposure, BHV-4 (strain FCAHV) was reisolated from explanted urinary bladder tissues of 5 of 6 exposed cats. Virus was not isolated from tissues of control cats. It was concluded that BHV-4 (strain FCAHV) establishes persistent urinary tract infection in conventionally reared adult male and female cats. However, persistent BHV-4 infection in cats may remain clinically inapparent.

A field strain (87-8363) of bovid herpesvirus-4 (BHV-4) isolated from an aborted bovine fetus was used to inoculate pregnant rabbits. Eleven rabbits in midgestation were alloted to 4 groups consisting of 3 infected groups and 1 control group. Rabbits were inoculated with BHV-4 or mock-infected cell culture preparations via IV, intravaginal, and intrauterine routes. Mild vulvovaginitis and endometritis were observed after intravaginal and IV inoculation of BHV-4, whereas intrauterine inoculation of BHV-4 resulted in abortion of hemorrhagic fetuses and nonsuppurative endometritis. Virus was successfully isolated from organ explants of fetal tissues. Rabbits seroconverted 1 week after infection as detected by results of an indirect immunofluorescence assay.

Serologic analyses and virus isolation studies were carried out to determine the role of bovid herpesvirus-4 (BHV-4) in infections in cattle, principally those of the reproductive tract. Serologic analyses were performed, using an indirect fluorescent antibody test on thoracic fluid specimens from aborted fetuses and on sera from 3 sources of adult cattle. Virus isolation was attempted from field cases of abortion, early embryo death, and postpartum vulvovaginitis/metritis, using uterine discharge and buffy coat preparations obtained from cows and tissues obtained from aborted fetuses. Of 420 fetal thoracic fluid specimens examined, 5 were positive for BHV-4 antibodies. Seventeen percent of adult cattle from 2 sources ie, clinically normal herds and abattoir cattle, were seropositive for BHV-4 antibodies. Cattle from a third source, 4 herds with high incidence of reproductive tract disorders, had a seroprevalence rate between 36 and 88%. Two isolates of BHV-4 were also obtained from this group. the overall incidence of BHV-4 antibodies in clinically normal cattle was higher than previously recognized, with relatively higher prevalence in herds having reproductive problems (chi-squared = 156.5, P less than 0.005). At least 10% of the BHV-4 antibody-positive sera did not have neutralizing antibody against bovine viral diarrhea virus and/or bovid herpesvirus-1, both important causes of bovine reproductive tract disorders.

A sensitive and specific DNA probe for detection and identification of bovine herpesvirus 4 (BHV-4) was developed. Cloned fragments from a library of HindIII fragments of the BHV-4 (DN-599) genome were labeled with 32P or digoxigenin and were tested for sentitivity and specificity in detecting viral DNA by dot-blot hybridization. Two probes were identified that detected 10 pg of purified viral DNA, and detected viral DNA in 0.001 microgram of total DNA extracted from BHV-4-infected cells. Both probes labeled with 32P and 1 labeled with digoxigenin detected viral DNA in samples prepared from cells infected with 2 prototype strains (DN-599 and Movar 33/63) and 4 field isolates of BHV-4. The DNA probes did not hybridize to total DNA prepared from uninfected bovine cells or from cells infected with BHV-1, BHV-2, alcelaphine herpesvirus 1, pseudorabies virus, or equine herpesvirus 1. One probe, labeled with digoxigenin, was tested further by dot-blot hybridization with infected cell lysates that were simply treated with sodium dodecyl sulfate and proteinase K prior to application to the membrane, avoiding extensive DNA purification procedures. This simplified procedure also resulted in specific detection of field isolates of BHV-4 and prototype strains of BHV-4.

Polymerase chain reaction was used to detect an economically important herpesvirus, channel catfish virus (CCV). A segment of the viral DNA was sequenced and oligonucleotide primers were produced from that sequence. After the primers were tested for the possibility of hybridization to catfish DNA, they were used to prime the polymerase chain reaction, using pure CCV DNA, CCV DNA added to catfish DNA, and DNA from catfish infected and not infected with CCV. In all cases, the method proved to be simple and sensitive in its detection of CCV DNA. When catfish DNA was present, < 0.1 pg of CCV DNA was detectable. Channel catfish virus DNA in a latent carrier of CCV was readily detectable.