Development of a Multiplex PCR Assay for Rapid Differentiation of Fowlpox and Pigeonpox Viruses.

The aim of this study was to develop a multiplex PCR assay capable of rapidly differentiating two major Avipoxvirus (APV) species, Fowlpox virus (FWPV) and Pigeonpox virus (PGPV), which cause disease in bird species. Despite the importance of a rapid differentiation assay, no such assay exists that can differentiate the APV species without sequencing. To achieve this, species-specific target DNA fragments were selected from the fpv122 gene of FWPV and the HM89_gp120 gene of PGPV, which are unique to each genome. Nine samples collected from unvaccinated chickens, pigeons, and a turkey with typical pox lesions were genetically identified as FWPV and PGPV. The designed primers and target DNA fragments were validated using in silico analyses with the nucleotide Basic Local Alignment Search Tool. The multiplex PCR assay consisted of species-specific primers and previously described PanAPV primers (genus-specific) and was able to differentiate FWPV and PGPV, consistent with the phylogenetic outputs. This study represents the first successful differentiation of FWPV and PGPV genomes using a conventional multiplex PCR test. This assay has the potential to facilitate the rapid diagnosis and control of APV infections.

Desarrollo de un ensayo de PCR múltiple para la diferenciación rápida de los virus de la viruela aviar y la viruela de paloma. El objetivo de este estudio fue desarrollar un ensayo de PCR múltiple capaz de diferenciar rápidamente dos especies principales de Avipoxvirus (APV) (viruela del pollo), el Fowlpox virus (FWPV) y el Pigeonpox virus (PGPV), (viruela de la gallina), que causan enfermedades en especies de aves. A pesar de la importancia de un ensayo de diferenciación rápida, no existe ningún ensayo que pueda diferenciar las especies de APV sin secuenciación. Para lograr esto, se seleccionaron fragmentos blanco de ADN específicos de especie del gene fpv122 de FWPV y el gene HM89_gp120 de Pigeonpox virus, que son únicos para cada genoma. Nueve muestras recolectadas de pollos, palomas y un pavo que no fueron vacunados con lesiones típicas de la viruela se identificaron genéticamente como FWPV y PGPV. Los iniciadores diseñados y los fragmentos de ADN blanco se validaron mediante análisis in silico mediante la herramienta de búsqueda de alineación local básica de nucleótidos (BLAST). El ensayo de PCR múltiple consistió en iniciadores específicos de especie y cebadores PanAPV previamente descritos (específicos de género) y fue capaz de diferenciar entre Fowlpox virus y Pigeonpox virus, de acuerdo con los resultados filogenéticos. Este estudio representa la primera diferenciación exitosa de los genomas de Fowlpox virus y Pigeonpox virus utilizando una prueba de PCR múltiple convencional. Este ensayo tiene el potencial de facilitar el diagnóstico rápido y el control de las infecciones por Avipoxvirus.

Encapsulation and drying methods in the production of powdered red cabbage (Brassica oleracea L.): Chemometrics and Fourier transform infrared spectroscopy.

In this study, powder colorant was obtained from red cabbage (Brassica oleracea L.). The stability of the colorants obtained by spray and freeze drying was investigated in terms of antioxidant capacity and anthocyanin content. The yield of the products increased with the encapsulation for both drying methods and encapsulation application. Drying method and encapsulation application had a significant effect on most of the physical properties of powders except for flowability and adhesiveness values. An increase in L*, a*, and C values was observed with the encapsulation process. Antioxidant activity of the samples increased with the encapsulation process by 13.44% in the spray-dried samples, while it increased by 9.75% in the freeze-dried samples. Total monomeric anthocyanin content was detected as 9039.21 mg/kg for encapsulated freeze-dried samples and 7811 mg/kg for encapsulated spray-dried ones. Nine anthocyanins were detected in the samples by using high-performance liquid chromatography. To discriminate samples according to drying methods with/without encapsulation principal component analysis (PCA) was used based on the Fourier transform infrared (FTIR) data. Four groups were observed for the PCA. The chemometric evaluation was done to predict the antioxidant capacity, anthocyanin content, and individual anthocyanins using FTIR spectra. High correlations were observed between the calculated and reference values for partial least square regression analysis.

Seroprevalence of Rhodococcus equi in horses in Israel.

Rhodococcus equi is a common cause of pneumonia in foals and has extensive clinical, economic and possibly zoonotic consequences. This bacterium survives well in the environment and may be considered as normal flora of adult horses. Certain strains of this bacterium are extremely virulent in foals, and early identification and intervention is crucial for prognosis. Rhodococcus equi is endemic in many parts of the world and occasionally isolated in Israel. This study was designed to evaluate R. equi seroprevalence in adult horses in Israel to indirectly indicate the potential level of exposure of susceptible foals. Sera were collected from 144 horses during spring 2011 and from 293 horses during fall 2014, and the presence of antibodies against virulent R. equi was detected by enzyme-linked immunosorbent assay. Equine seroprevalence of R. equi was found to be 7.6% in 2011 and 5.1% in 2014. Only one farm had seropositive horses in 2011, whereas several farms had seropositive horses in 2014. No significant risk factors for seropositivity were found. Rhodococcus equi appears to be endemic in Israel. This is the first survey of R. equi in Israel that provides information on the epidemiology of this important bacterium.

Salmonella, Campylobacter, and Chlamydophila in bald ibis (Geronticus eremita) feces in Turkey.

The aim of this study was to investigate the presence of Campylobacter spp., Salmonella spp., and Chlamydophila psittaci in fecal samples of bald ibises (Geronticus eremita) housed in a conservation facility in Turkey. A total of 82 fecal samples were collected from cages and evaluated by bacteriologic methods and a polymerase chain reaction (PCR) technique for Campylobacter spp. and Salmonella spp. and by PCR for C. psittaci. Campylobacter spp. were isolated from 24 of 82 fecal samples (29.2%). Of these 18 (75%), 4 (16.7%) and 2 (8.3%) were Campylobacter jejuni, Campylobacter coli, and other Campylobacter spp., respectively. Salmonella spp. were detected in 8 fecal specimens.(9.7%) by PCR. The presence of C. psittaci was not detected in the bald ibises studied. The results suggested that the bald ibises in this present study might be at a higher risk of infection with Salmonella spp. and Campylobacter spp.

Investigation of the antibiotic resistance and biofilm formation of Staphylococcus aureus strains isolated from gangrenous mastitis of ewes.

In this study, Staphylococcus aureus strains (n = 110) isolated from seven ewe flocks in Sanliurfa, Turkey were screened for antibiotic resistance and biofilmforming ability as well as for genes associated with antibiotic resistance and biofilm-forming ability. All isolates were found to be susceptible to oxacillin, gentamicin, clindamycin, cefoxitin, tetracycline, vancomycin, amoxicillin-clavulanic acid, ciprofloxacin and sulphamethoxazole-trimethoprim. The percent proportions of strains resistant to penicillin G, ampicillin and erythromycin were 27.2% (n = 30), 25.4% (n = 28) and 6.3% (n = 7), respectively. Regarding the antibiotic resistance genes, 32 (29%) isolates carried the blaZ and 8 (7.2%) the ermC gene. Other resistance genes were not detected in the isolates. All isolates showed biofilm-forming ability on Congo red agar (CRA), while 108 (98.18%) and 101 (91.81%) of them were identified as biofilm producers by the use of standard tube (ST) and microplate (MP) methods, respectively. All isolates carried the icaA and icaD genes but none of them harboured the bap gene. The results demonstrated that S. aureus isolates from gangrenous mastitis were mainly resistant to penicillins (which are susceptible to the staphylococcal beta-lactamase enzyme), and less frequently to erythromycin. Furthermore, all of the S. aureus isolates produced biofilm which was considered a potential virulence factor in the pathogenesis of staphylococcal mastitis.