Validation of RT‐PCR Assays for Molecular Characterization of Porcine Teschoviruses and Enteroviruses

Porcine enteroviruses (PEVs) and teschoviruses (PTVs) are described as causative agents of neurological disorders, fertility disorders and dermal lesions of swine. Difficulties in the serological detection of these viruses may lead to a significant underestimation of infections with clinical symptoms. With the recent availability of genome sequence data for all the serotypes, molecular diagnosis is a possibility. The present study describes a new approach to molecular ‘serotyping’ of PTVs and PEV‐B viruses, involving the amplification and sequencing of a genomic fragment of the VP1 coding region. A molecular characterization of Italian entero‐teschovirus isolates was performed using a set of previously published and newly designed polymerase chain reaction primers. A total of 33 porcine isolates and 10 reference strains were analysed. Porcine enterovirus‐B samples were first diagnosed as positive for enterovirus by amplification of the 5′‐non‐translated region. Samples were then typed by amplification and sequencing of a portion of the VP1 coding region. Porcine enterovirus‐A and PTVs were detected by a published assay in the 5′‐NC region that allows them to be differentiated according to the size of amplification product, using the same set of primers. For serotype characterization of PTV, we evaluated four different regions: the N terminus of the capsid protein VP2, the region encoding for RNA‐dependent RNA polymerase, and the capsid VP1 and VP4 regions. The newly designed primers in the VP1 region was proved to be broad in range and suitable for serotype assessment and therefore constitute a useful diagnostic tool for molecular diagnosis of porcine teschovirus/enterovirus strains and for the study of molecular epidemiology and evolution of these viruses.     

Socio-spatial organization reveals paternity and low kinship in the Honduran white bat (Ectophylla alba) in Costa Rica

Ectophylla alba is a tent-making bat that roosts in mixed-sex clusters comprising adults and offspring. Our goal was to determine the genetic identity of individuals belonging to different roosting groups. We tested the hypothesis of kin selection as a major force structuring group composition. We used 9 microsatellites designed for E. alba to determine the genetic identity and probability of parentage of individuals. We analyzed parentage and kinship using the software ML-Relate, GenAIEx, and Cervus. The obtained relationship probabilities (0.5) revealed a clear maternal relationship between female adults and offspring with allele compatibility, and at least 5 relationships between male adults and pups. We found a low degree of relatedness within roosting groups. Between roosting groups at different sites, the mean probability of a half-sibling relationship ranged from 0.214 to 0.244 and, for full-sibling relationship, from 0.383 to 0.553. Genetically, adult individuals were poorly related within clusters, and kinship as an evolutionary force could not explain group membership.     

Efficacy of a single milbemycin oxime administration in combination with praziquantel against experimentally induced heartworm (Dirofilaria immitis) infection in cats

The efficacy of a combination of milbemycin oxime and praziquantel in preventing the establishment of experimentally induced heartworm (Dirofilaria immitis) infection was investigated in a study involving 24 young domestic short-hair cats. The animals were inoculated with 50 infective larvae on day 0. Subsequently they were divided into two groups of 12 animals each. The animals in group 1 were treated once with medicated tablets containing 4 mg milbemycin (minimum dose 2 mg/kg body weight) and 10 mg praziquantel (MILBEMAX) on day 30 after infection. Cats in group 2 received placebo tablets on the same day. On day 183 post-infection a blood sample was taken from each animal before euthanasia and necropsy. The blood samples were tested for the presence of microfilariae and the necropsied animals were examined for the presence of adult worms. Microfilariae were not found in any of the investigated cats. No heartworms were found in the animals in group 1 (treated with medicated tablets). Out of the 12 placebo-treated cats 1 was heartworm-free, whereas all the others were found to be infected with 1-3 adult heartworms.     

Evaluation of head-out constant volume body plethysmography for measurement of specific airway resistance in conscious, sedated sheep

OBJECTIVE: To evaluate the use of a modified whole body plethysmograph in awake sheep. ANIMALS: 10 healthy adult sheep. PROCEDURE: Concurrent measurements of specific airway resistance (sR(aw)) and pulmonary resistance (R(L)) were obtained using a novel noninvasive head-out constant-volume plethysmograph and esophageal balloon-pneumotachography, respectively. All data were collected before and after external resistive loading with 1 and 5.6 cm H2O/L/s. Functional residual capacity (FRC) was measured by helium dilution for computation of airway resistance (R(aw)) preloading (R(aw) = sR(aw)/FRC). RESULTS: The sR(aw) and R(L) were closely correlated in 10 adult sheep. Additionally, sR(aw), and R(L) accurately reflected the magnitude of added resistance. The mean FRC was 52 mL/kg and used to calculate R(aw). At baseline, the values for R(aw) were significantly correlated with sR(aw) and R(L). CONCLUSIONS AND CLINICAL RELEVANCE: Precise measurements of sR(aw) and R(aw) at baseline and sR(aw) after external resistive loading were obtained by use of this novel noninvasive plethysmographic technology. This method should have application to veterinary patients or animals used in research in which noninvasive rapid or serial measurements of sR(aw) in the conscious state are required.     

Long‐term residence of juvenile loggerhead turtles to foraging grounds: a potential conservation hotspot in the Mediterranean

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Extracts of the endemic resurrection plant Haberlea rhodopensis stimulate in vitro growth of various Phytophthora spp. pathogens

The Phytophthora pathogens are among the most important pests in the modern agriculture and forestry. This makes them a subject of intensive studies, including laboratory experiments with in vitro culture on various plant-derived media. Here we show that leaf extracts from the resurrection endemic plant Haberlea rhodopensis stimulates significantly the in vitro growth of various isolates from several Phytophthora species. The extracts stimulate the formation of oospores in some heterothallic species. In this respect, the idea to propose Haberlea extracts as a potential ingredient of culture media for Phytophthora spp. maintenance under controlled conditions or as a component of detection systems for Phytophthora presence in nature seems quite attractive and promising.     

MALDI‐MS Lipid Profiles of Oocytes Recovered by Ovum Pickup from Bos indicus and 1/2 indicus × taurus with High vs Low Oocyte Yields

The aim of the present study was to compare the lipid profile in oocytes of indicus and 1/2 indicus × taurus cows with high and low antral follicle count (AFC)/oocyte yields. After an OPU procedure (D0), antral follicles ≥3 mm were counted by ultrasonography (D4, 19, 34, 49, 64), and cows were assigned to groups with either high AFC (≥30 follicles; indicus, NH group; 1/2 indicus × taurus, AH group) or low AFC (≤15 antral follicles; indicus, NL group; 1/2 indicus × taurus, AL group). The lipid profiles of the oocytes were determined by MALDI‐MS. For GI, GII and GIII oocytes, the indicus samples tend to cluster separately from the 1/2 indicus × taurus samples. The lipid species [PC (P‐38:5) + H]⁺ and/or [PC (P‐36:2) + Na]⁺, [PC (38:2) + H]⁺, [PC (38:5) + Na]⁺ and [TAG (60:8) + NH⁴]⁺ were more abundant in indicus (NH and NL groups) than 1/2 indicus × taurus. The higher lipid content in the indicus oocytes likely reflects differences in the rate of lipid metabolism and may contribute to oocyte competence and embryo development.     

Vaccination of one-day-old broiler chicks against infectious coryza

In order to evaluate the protection conferred by an experimental inactivated vaccine against infectious coryza, three challenge trials were undertaken using 112 1-day-old broilers. The vaccine "Hepa Inmuno NC" included bacterial antigens of Avibacterium paragallinarum (serogroups A, B, variant B, and C) as well as antigens of Newcastle virus and hepatitis virus. Fifty-six broiler chicks were vaccinated at the first day of life at the hatchery while another 56 chicks were left unvaccinated. Three infection trials were conducted simultaneously using each of the three serogroups A, B, or C of Av. paragallinarum. In each trial, 17 vaccinated and 17 unvaccinated broilers were used. Challenge was performed at day 31 of life by injection, into the left infraorbital sinus, of approximately 1 x 10(5) colony forming units of the corresponding Av. paragallinarum strain. Clinical signs were recorded on day 2 postchallenge. All broilers were euthanatized and both infraorbital sinuses were bacteriologically examined for the presence of Av. paragallinarum on day 5 postchallenge. In comparison with the unvaccinated broilers, the vaccine significantly reduced the number of broilers with clinical signs after challenge with serogroup B, and significantly fewer vaccinated broilers were positive for the presence of Av. paragallinarum after challenge with serogroup C. On the other hand, no significant protection was observed when broilers were challenged with Av. paragallinarum from serogroup A. Despite the high infection rates in vaccinated chicks after an experimental infection with Av. paragallinarum, it was possible to reduce colonization of Av. paragallinarum (serogroup B) and clinical signs (serogroup C) in broiler chicks by vaccination at the first day of life. Further cross-protection trials should be done, including other Av. paragallinarum strains in the vaccine, especially those from serogroup A.     

Comparison of Regular and Selenium-Enriched Tortillas Produced from Sprouted Corn Kernels

Plant Foods for Human Nutrition - Corn kernels were soaked with different selenium (Se) solutions (0, 12 or 24 mg Na2SeO3/L), sprouted for different times and then lime-cooked for the...