FC‐14 Clonality studies of feline cutaneous lymphocytosis

A previous study described cutaneous lymphocytosis (CL) in 23 cats. The process resembles cutaneous pseudolymphoma in humans, a heterogeneous group of benign reactive proliferations of well‐differentiated lymphocytes in the skin of humans. Morphological and immunophenotypic characteristics do not offer reliable criteria to accurately predict the clinical outcome of feline CL or pseudolymphoma in humans. Presence of clonal cell populations is more consistent with a neoplastic process. In a previous study, feline CL lesions (20 cats) were evaluated for clonality using PCR, and only two cats had monoclonal T‐cell populations. Because false‐negative results may occur, the purpose of this study was to repeat the PCR using a revised primer set based on analysis of additional feline T‐cell receptor γ (TCRγ) sequences. DNA was isolated from 29 skin lesions and six internal organs of 20 cats. DNA integrity was assessed by glyceraldehyde‐3‐phosphate dehydrogenase PCR. Polymerase chain reaction clonality was performed using the revised primer set specific for feline TCRγ, and duplicate samples were evaluated. The PCR products were assessed by heteroduplex analysis. Clonal rearrangement of TCRγ was detected in 14 cats (24 of 35 tissues: 21 of 29 skin lesions and three of six internal organs); eight of these cats are still alive and six were euthanized. Monoclonal populations were seen in three of five cats that had involvement of internal organs. These findings indicate that feline CL is best considered as a slowly progressive process which may be reactive, but often evolves into a low‐grade indolent lymphoma. Funding: George H. Muller Fund for Research in Dermatology.     

Dirofilaria repens in a cat with acute liver failure

Acute liver failure was diagnosed in a 12-year-old cat. Fine needle aspirate cytology revealed high numbers of unsheathed microfilariae and a hepatocellular reaction with no evidence of bacterial infection. The microfilariae were identified as those of Dirofilaria repens by acid phosphatase staining. The high number of microfilariae seen in both the blood and the liver aspirate samples as well as the favourable response to ivermectin amongst other drugs administered, is suggestive that D. repens was the cause of the liver insult. A positive result obtained with an antigen-capture ELISA (Dirochek) for Dirofilaria immitis antigen was interpreted as false. This is the 1st report of Dirofilaria repens for South Africa.     

Effects of porcine reproductive and respiratory syndrome virus (isolate tw91) on porcine alveolar macrophages in vitro

To verify the role of porcine reproductive and respiratory syndrome virus (PRRSV) infection on pulmonary defense mechanisms, alterations in the viability, morphology, and various functions of porcine alveolar macrophages (AMs) were evaluated in vitro for 2-72 h after exposure to a Taiwan isolate, tw91, at a multiplicity of infection (MOI) of 0.1. A low but constant rate of infection, around 5%, was seen in AMs from the PRRSV-infected group throughout the study. When compared with a mock-infected group, AMs from the PRRSV-infected group had a significantly lower viability at 18-72 h post-infection (HPI) as determined by trypan blue dye exclusion. Also during this time period, the cells showed morphological changes, including rounding, bleb formation, and rupture. The phagocytic and microbicidal capacity of AMs against Candida albicans was significantly inhibited after 6 HPI. Although the total amount of superoxide anion (O2-) and hydrogen peroxide (H2O2) produced by the AMs was reduced after 18 and 12 HPI, respectively, the amount of production was enhanced in both reactive oxygen species on a per viable cell basis after 12 HPI. In contrast, the level of bioactive tumor necrosis factor alpha (TNF-alpha) secretion, either total or on a per viable cell basis, was markedly reduced soon after PRRSV infection, up to 36 HPI, followed by a rebound thereafter. Prostaglandin E2 (PGE2) production was enhanced, both in total and on a per viable cell basis, in the first 6 h of infection, especially at 2 HPI. However, it became lower than that of the control after 36 HPI. The results indicated that PRRSV infection could cause, directly and/or indirectly, not only death of AMs but also adverse alterations in their morphology and function, although some of the effects seemed to be reversible. Because AMs are crucial to the host against airborne pathogens, PRRSV infection may potentially predispose pigs to secondary pulmonary infections.     

Relationships of sire scrotal circumference to offspring reproduction and growth

Reproductive and growth data were obtained on 779 and 564 yearling beef heifers and bulls, respectively, that had sires with yearling scrotal circumference data at the San Juan Basin Research Center, Hesperus, CO. Partial regression coefficients of reproductive and growth traits on inbreeding (FXC) and age of the individual and adjusted scrotal circumference of sire (SCSI) were obtained. Growth and reproductive traits of heifers and growth and breeding soundness traits of bulls were analyzed. Separate analyses for each sex were performed, but least squares models were similar. Models included fixed effects of breed, birth year (BY), age of dam (AOD) and the covariates FXC, age (day of birth in heifer analyses) and SCSI. Scrotal circumference of sire was adjusted for age, FXC, AOD and BY using values obtained in a separate analysis. Seminal traits improved as age increased, and there was a seasonal effect present for age of puberty. Inbreeding had a detrimental effect on reproductive traits. Partial regression coefficients for the reproductive traits on SCSI were: age of puberty, -.796 d/cm; age of first calving, -.826 d/cm; julian day of first calving, -.667 d/cm; julian day of second calving, .597 d/cm; most probable producing ability, .132 %/cm; percent sperm motility, -.74 %/cm; percent primary sperm abnormalities, .08 %/cm; percent secondary sperm abnormalities, .92 %/cm; percent normal sperm, -1.28 %/cm; total breeding soundness examination score, .28 units/cm and scrotal circumference, .306 cm/cm. A heritability of .39 was obtained for scrotal circumference.     

First Isolation of Rotavirus Associated with Neonatal Diarrhoea in Guanacos (Lama guanicoe) in the Argentinean Patagonia Region

Group A rotavirus (RV) and coronavirus (CV) are common viral pathogens associated with neonatal diarrhoea in numerous animal species. The purpose of this work was to investigate the presence of these viral agents in two farm populations of captured guanacos (Lama guanicoe) in the Argentinean Patagonia region, that developed severe diarrhoea outbreaks. Stool and serum samples were analysed for RV and bovine CV antigen and antibody by enzyme‐linked immunosorbent assay. Rotavirus was detected in faeces from two new‐born guanacos with acute diarrhoea, one in each farm. After electrophoretic analysis, each isolated strain, showed a distinctive long dsRNA electropherotype characteristic of group A rotaviruses (4:2:3:2). In addition, 95% (38 of 40) of the sampled animals were positive for RV antibodies, suggesting a high prevalence of RV infection in the populations tested. No evidence of CV circulation by antigen or antibody analysis was observed. To our knowledge, this is the first report of the detection and isolation of RV associated with neonatal diarrhoea in Lama guanicoe.     

Identification,pathogenicity and community dynamics of fungi and oomycetes associated with pea root rot in northern France

The pea root rot complex is a major concern for green pea production worldwide. This study aimed at characterizing its composition and dynamics throughout a cropping season in northern France. To this end, fungi and oomycetes were isolated from green pea plant roots with symptoms sampled at the flowering stage in 22 fields in 2017, and at the pea emergence, elongation and flowering stages in two fields in 2018. Out of 646 isolates collected, 317 were identified using molecular markers. Fusarium oxysporum, F. solani and F. redolens were highly predominant. Pathogenicity tests separated the isolates into four aggressiveness groups. F. solani isolates were the most aggressive. Phylogenetic analysis of their TEF1 sequences showed that they mainly belonged to the F. pisi lineage, and that F. oxysporum isolates were genetically close to isolates from the UK that did not belong to the forma specialis pisi. In addition, several Clonostachys rhizophaga isolates are reported for the first time to cause pea root rot. The oomycetes were rarely found and were represented by a few Pythium spp. isolates. Lastly, this study shows that the fungal and oomycete communities associated with pea root rot change during the cropping season. The level of dissimilarity of the root-rot-associated communities decreased throughout the cropping season towards a more similar composition at the flowering stage, dominated by F. solani, F. oxysporum and F. redolens. The proportion of nonpathogenic to weakly pathogenic isolates decreased progressively during the growing season in favour of moderately to highly pathogenic isolates.     

Variation at the b Mating Type Locus of Ustilago maydis

ABSTRACT Population level diversity at the Ustilago maydis b mating type locus was determined in samples from four Minnesota locations using a combination of plate mating techniques and a polymerase chain reaction (PCR)-based assay. The PCR method allows rapid identification of b types from samples of natural populations and utilizes the hypervariable regions of the b locus that determine mating type specificity. Results demonstrated high levels of b diversity within populations, with one population yielding 17 of the total 18 b types found in the study. Pairwise G(ST) values were in the range of 0.02 to 0.05, and common b mating types were found across broad geographic distances. These data demonstrated that very low levels of differentiation among U. maydis populations occur with respect to b locus variation. Consistent with frequency-dependent selection models, b types were represented at approximately equal frequencies within the entire Minnesota population. However, neutral evolutionary models for patterns of geographic distribution and variation at b cannot be entirely excluded. The importance to agricultural practices of understanding population genetic processes is discussed.     

Planting density for grafted melon as an alternative to methyl bromide use in Mexico

One of the major challenges of melon (Cucumis melo L.) yield is the decrease of fruit yield and quality caused by soil diseases. Soil disinfection with methyl bromide (MB) has been used to prevent soil-borne diseases damage; however, use of MB is being restricted because this substance damages the ozone layer. Searching for new MB alternatives, field experiments were carried out in open fields in soils infested with Fusarium oxysporum f. sp. melonis, Olpidium bornovanus, and Melon Necrotic Spot Virus (MNSV) in Colima, Mexico, where melons had only been grown previously using soil fumigation with MB. Yield and quality of melon cultivar Cruiser F1 (Harris Moran seed company) grafted on two rootstocks of Cucurbita maxima Duchesne × Cucurbita moschata Duchesne (“RS841”; Séminis seed company and “Shintosa Camelforce”; Nunhens seed company) were evaluated during two consecutive years. Each experiment had five treatments, two of which with non-grafted plants at a density of 18,519 plants/ha, with and without MB fumigation. The other three treatments consisted of grafted plants in non-fumigated soil with plant densities of 14,815, 11,111, and 9260 plants/ha. The use of melon grafted on “RS841” and “Shintosa Camelforce” rootstocks significantly increased the average fruit weight and the total yield of melons in soil without the need for MB. Grafting may be considered an alternative to MB fumigation. With the use of grafted melon plants, planting density may be reduced by 60%, obtaining higher yields than those obtained from non-grafted plants grown on fumigated land. Fruits harvested from grafted plants had equal firmness than those harvested from non-grafted plants, without affecting the content of soluble solids.