Complete Genome Sequence of an Ehrlichia minasensis Strain Isolated from Cattle

Ehrlichia minasensis is a tick-borne pathogen affecting cattle, cervids, and dogs, and it is closely related to the monocytotropic pathogen Ehrlichia canis. Here, we announce the draft genome sequence of Ehrlichia minasensis strain Cuiabá, isolated from a naturally infected calf from Santo Antônio do Leverger, Mato Grosso, Brazil.

Complete Genome Sequence of Bovine Viral Diarrhea Virus Subgenotype 2a Strain CN10.2015.821, Isolated in Piedmont, Italy

ABSTRACT We sequenced the complete genome of noncytopathic bovine viral diarrhea virus (BVDV) type 2 strain CN10.2015.821. It belongs to the subgenotype 2a, and it was isolated from an immunotolerant and persistently infected calf identified during a serological investigation. The complete genome is composed of 12,273 nucleotides, organized as one open reading frame encoding 3,897 amino acids.

Border Disease Virus Infection of Bovine Placentas

Subsequent to a previous study of border disease virus (BDV) horizontal transmission from a persistently BDV-infected calf to 6 seronegative pregnant heifers, the heifers were slaughtered 60 days after exposure to the infected calf, and their fetuses and placentas were examined. Immunohistochemical examination of fetal organs and placenta showed positive labeling of moderate intensity for pestivirus antigen in 3 of 6 heifers. BDV infection in these 3 animals was confirmed by the detection of BDV RNA in different organs using reverse transcription quantitative polymerase chain reaction. In the placenta, the positive cells were visualized mostly on the fetal side. In those 3 heifers that harbored an infected fetus, the placental tissue in the placentome region showed a moderate to severe mononuclear and fibrosing placentitis and, in severe cases, necrotic areas. The inflammatory population was composed predominantly of T and B cells, a substantial number of macrophages, and, to a lesser extent, plasma cells. This is a novel report of placentitis in persistently BDV-infected fetuses from pregnant heifers that became acutely infected by cohousing with a calf persistently infected with BDV, which extends previous reports on bovine viral diarrhea virus–infected and BDV-infected cattle and sheep, respectively.

On the Efficacy and Safety of Vaccination with Live Tachyzoites of Neospora caninum for Prevention of Neospora-Associated Fetal Loss in Cattle

ABSTRACTInfection of cattle withNeospora caninummay result in abortion or the birth of a congenitally infected calf. Vaccination with liveN. caninumprotects against experimental infection of cattle and mice, and the naturally attenuated Nc-Nowra strain ofN. caninumis of particular interest as a potential vaccine candidate. Vaccination of heifers prior to breeding with live Nc-Nowra tachyzoites by either the subcutaneous or the intravenous route reduced the rate of abortion and the presence of the parasite in calves as determined by PCR and serology after infection of cows with a virulent isolate. Protected fractions were 55.6% to 85.2% depending on the route of vaccination and growth conditions of the vaccine strain, with cryopreserved Nc-Nowra tachyzoites being less effective, with a 25.9% protected fraction. Vaccination appeared to reduce the rate of pregnancy after artificial insemination in some groups compared to nonvaccinated, nonchallenged controls. One animal that was vaccinated but not challenged experienced an abortion, but Nc-Nowra could not be detected in any of the cows in this group or their progeny. This study confirms that live vaccination can be an effective method of preventing neosporosis in cattle and yet highlights the technical hurdle of preservation of live parasites that must be overcome for a vaccine to be commercially successful.

Isolation and biological characterisation of a new isolate of Neospora caninum from an asymptomatic calf in Brazil

Neospora caninum is a tissue-cyst forming parasite that has been recognized worldwide as a cause of abortion in cattle. Despite the ubiquitous distribution of this parasite and its broad range of hosts, the number of N. caninum isolates obtained to date is limited. In addition, the majority of isolates have been obtained from clinically affected hosts, therefore potentially biasing this population towards more virulent isolates. This report describes the isolation and biological characterisation of a new N. caninum isolate, Nc-Goiás 1, obtained from an asymptomatic, naturally infected calf from Brazil. This new isolate was identified as a member of the N. caninum species by polymerase chain reaction (PCR) using specific primers based on the N. caninum internal transcribed spacer 1 (ITS-1) sequence, and was genetically identified at multiple loci using microsatellite analysis. Finally, a pathogenicity study was conducted in a BALB/c mice model. All Nc-Goiás 1-infected mice survived without exhibiting any clinical signs. Further pathogenic characterisation of this isolate suggested that Nc-Goiás 1 is less virulent than other N. caninum isolates (Nc-Liv and Nc-1) studied in this mouse model. This is the first report of the isolation and biological characterisation of N. caninum from an infected but clinically healthy calf in South America.

Heterologous transmission with Dictyocaulus capreolus from roe deer (Capreolus capreolus) to cattle (Bos taurus)

Eight Swedish Red Breed cattle, about 2 months old, were experimentally infected with a Swedish isolate of Dictyocaulus viviparus (Dviv-Se) from cattle and D. capreolus from roe deer. The aims were to determine whether the roe deer lungworm is infective to cattle or if it can induce seroconversion in cattle against D. viviparus as measured with an ELISA. Four calves which were given 500 Dviv-Se infective larvae (L3) each by larval dosing for two successive days developed patent infection between days 23 and 25 post-inoculation (PI). Larval output varied among the calves and during the patent period. However, maximum recovery occurred between 28 and 56 days PI with peak shedding on day 37 PI. Shedding ceased at day 58 PI and adult worms were recovered from one calf at necropsy (day 67 PI). No immature worms were recovered from the lungs at necropsy. Seroconversion was detected on days 35–42 PI. One Dviv-Se infected calf became seronegative on day 67 PI whereas the other calves still remained seropositive during this period. Prepatency and patency periods of D. viviparus and serological findings in this study basically conform to previous studies. Each calf that was infected with 400 L3 of D. capreolus for two successive days, and about 800 L3 of the same species about 8 weeks later, did not develop to patency based on faecal and post-mortem examinations. Consequently, under the conditions of this study, D. capreolus was not infective to cattle. Two of the four calves that were infected with L3 from roe deer were challenged with L3 cultured from faeces of the Dviv-Se-infected calves. This infection did not develop to patency. Whether this was due to cross-protection as a result of the prior priming with L3 from roe deer is not clear. However, if it is so, it opens up the possibility of using D. capreolus L3 for preventing bovine dictyocauliasis.

Neospora caninum-associated abortion in cattle: the time of experimentally-induced parasitaemia during gestation determines foetal survival

The parasite, Neospora caninum is an important cause of abortion in cattle. It is transmitted vertically or horizontally and infection may result in abortion or the birth of a live, healthy but infected calf at full-term. Only a proportion of infected cattle abort and the pathogenesis of abortion is not understood. Groups of cattle were infected with 107N. caninum tachyzoites intravenously at different times relative to gestation. Intravenous inoculation was chosen to reproduce the putative haematogenous spread of N. caninum following either recrudescence of endogenous infection or de novo infection. In all cattle, infection was accompanied by high γ-interferon and lymphoproliferative responses, and a biased IgG2 response indicating that N. caninum infection is accompanied by a profound Th1 helper T cell-like response. Infection at 10 weeks gestation resulted in foetopathy and resorption of foetal tissues 3 weeks after infection in 5 out of 6 cows. Infection at 30 weeks gestation resulted in the birth of asymptomatic, congenitally-infected calves at full term in all 6 cows, whereas the 6 cows infected before artificial insemination gave birth to live, uninfected calves. These results suggest that the reason some cows abort is related to the time during gestation when they become infected or an existing infection recrudesces.

Inactivation of Cryptosporidium parvumOocysts by Ammonia

ABSTRACT The survival of Cryptosporidium parvum oocysts in soil and water microhabitats may be affected by the environmental production and release of free ammonia. The objective of this study was to determine the effects of increasing free ammonia concentrations and times of exposure on oocyst viability. Wild-type oocysts were obtained from naturally infected calf feces by chemical (continuous-flow) centrifugation and sucrose gradients. Ammonia (NH3) from a commercial solution was applied in concentrations ranging from 0.007 to 0.148 M. Exposure times ranged from 10 min to 24 h at a constant temperature of 24 ± 1°C. Viability of oocysts was determined with a dye permeability assay and an in vitro excystation assay (M. B. Jenkins, L. J. Anguish, D. D. Bowman, M. J. Walker, and W. C. Ghiorse, Appl. Environ. Microbiol. 63:3844–3850, 1997). Even the lowest concentration of ammonia decreased significantly the viability of oocysts after 24 h of exposure. Increasing concentrations of ammonia increased inactivation rates, which ranged from 0.014 to 0.066 h−1. At the highest concentration of ammonia, a small fraction of viable oocysts still remained. Exposure to pH levels corresponding to those associated with the ammonia concentrations showed minimal effects of alkaline pH alone on oocyst viability. This study shows that environmentally relevant concentrations of free ammonia may significantly increase the inactivation of oocysts in ammonia-containing environments.