BACKGROUND: Coronaviruses have a wide range of host tropism causing respiratory, enteric and central nervous system diseases in pigs, cats, dogs, rodents, cattle, avian species and human. Coronaviruses undergo genetic mutations and recombination at high rates which make them able to infect a wide range of host species from different geographical locations. According to the possible existence of gammacoronavirus in Iranian bird population including pigeons and lack of information about virus prevalence, isolation and molecular characterization of pigeon coronaviruses are needed. OBJECTIVES: The present study was conducted to detect avian coronavirus in flocks of Tehran province.               METHODS: samples were randomly collected from 25 pigeon flocks of Tehran province (250 tracheal swabs and 250 cloacal swabs, 20 specimens from each flock) between 2014-2015. The viral RNA was extracted from swab samples and RT-PCR reaction was run using the QIAGEN one-step RT-PCR Kit with primers targeting nucleocapsid (N) gene and 3’ untranslated region (3’-UTR) of gammacoronavirus. RESULTS: Gammacoronavirus was detected in one out of 25 (4%) flocks. CONCLUSIONS: The results of this study approve the presence of  gammacoronaviruses  in pigeon population and help to complete the map of  epidemiology of the virus in Iran. According to the low prevalence rate of coronavirus in pigeons, samples should be collected from pigeons showing respiratory or enteric signs of disease or from pigeons having contact with other birds or those which are housed near poultry farms.

The aim of the study was to determine the effect of the vitamins, omega-3 polyunsaturated fatty acid and minerals in the supplement Toryum administered before and during oestrus synchronisation on some fertility parameters of ewes during the non-breeding season. The experimental animals were clinically healthy Pirlak ewes, 55–75 days postpartum, aged 2–4 years and weighing 40–50 kg. A sponge was inserted into the vagina for 10 d (G1, n = 30; G2, n = 30) or 14 d (G3, n = 30; G4, n = 30) for oestrus synchronisation, and on the day of removal, 400 IU equine chorionic gonadotropin was injected. Toryum soft capsules were administered individually (1 capsule/ewe p.o.) to G1 and G3 ewes seven days before the sponge was inserted and on the day it was removed. Oestrus detection was started 12 h after sponge removal. Pregnancy was diagnosed by transrectal ultrasonography on the 30ᵗʰ day after mating. The pregnancy rate was statistically different between G1 and G4 (P < 0.05). The onset of oestrus was statistically different (P < 0.001) between the 10-d groups (G1 and G2) and the 14-d groups (G3 and G4). The litter size and oestrus, conception, lambing, multiple birth, and survival rates were not significantly different between the groups (P > 0.05). Toryum administered to Pirlak ewes during progesterone-based oestrus synchronisation protocols during the non-breeding season may increase pregnancy rates. The relationship between Toryum and fertility parameters in ewes would be better understood by comprehensive studies.

The plate counting method widely used at present to discern viable from non-viable Brucella in the host or cell is time-consuming and laborious. Therefore, it is necessary to establish a rapid, simple method for detecting and counting viable Brucella organisms. Using propidium monoazide (PMA) to inhibit amplification of DNA from dead Brucella, a novel, rapid PMA-quantitative PCR (PMA-qPCR) detection method for counting viable Brucella was established. The standard recombinant plasmid with the target BCSP31 gene fragment inserted was constructed for drawing a standard curve. The reaction conditions were optimised, and the sensitivity, specificity, and repeatability were analysed. The optimal exposure time and working concentration of PMA were 10 min and 15 μg/mL, respectively. The correlation coefficient (R²) of the standard curve was 0.999. The sensitivity of the method was 10³ CFU/mL, moreover, its specificity and repeatability also met the requirements. The concentration of B. suis measured by the PMA-qPCR did not differ significantly from that measured by the plate counting method, and the concentrations of viable bacteria in infected cells determined by the two methods were of the same order of magnitude. In this study, a rapid and simple PMA-qPCR counting method for viable Brucella was established, which will facilitate related research.

White sturgeon iridovirus (WSIV) disease is caused by a virus of the eponymous family and is mostly triggered by stressful environmental conditions, i.e. high rearing density, excessive handling, or temporary loss of water. The aim of this study was to develop the most effective diagnostic method for quick and efficient confirmation or exclusion of the presence of WSIV. A total of 42 samples (spleen, gills, intestine, skin, kidney, and brain) were collected from eight sturgeon (Acipenser gueldenstaedtii and A. oxyrinchus) aged ≤5+ farmed or caught between 2010 and 2014 in open waters (Dąbie Lake and Szczecin Lagoon). They were tested for WSIV presence using conventional PCR, qPCR, and in situ hybridisation (ISH). In gross examination, all fish appeared to be healthy. Neither species showed clinical signs typical of WSIV infection. In the majority of cases, fragments of iridoviral DNA were found using molecular methods in the kidneys, and also in the liver, gills, and skin. The detection rate using ISH was 47.37% and most commonly the brain and kidney tissues were positive. The most efficient of the methods used was real-time PCR, with 100% effectiveness in detection of WSIV DNA. The study demonstrates the capabilities for WSIV diagnosis available to sturgeon farmers and water administrators, indicating useful methods of adequate sensitivity as well as organs to sample in order to achieve the highest probability of viral detection.

The aim of the study was to estimate the diagnostic sensitivity (DSe) and specificity (DSp) of an agar gel immunodiffusion (AGID) assay for detection of myxoma virus (MYXV) in the classical form of myxomatosis and to compare its diagnostic performance to that of molecular methods (IAC-PCR, OIE PCR, and OIE real-time PCR).

The aim of the study was to determine the transmission potential of carp edema virus (CEV) and koi herpesvirus (KHV) introduced to Europe by the invasive round goby (Neogobius melanostomus). A total of 70 round goby specimens were collected from the Szczecin Lagoon, Poland, and locations in Germany in the third and fourth quarters of 2018. The fish were analysed to detect KHV and CEV by PCR. Six fish specimens were positive for the presence of KHV, while none of the gobies examined showed the presence of CEV. The CEV genome was detected in the goby specimens from Germany and from Poland. Considering the high pace of the spread of the round goby and its effectiveness in acquisition of new ecological niches, it should be kept out during refilling of carp ponds. Further studies should focus on experimental cohabitation of CEV-infected round gobies and specific-pathogen-free (SPF) carp to investigate the potential for active virus transfer.

The aim of the study was to estimate the diagnostic sensitivity (DSe) and specificity (DSp) of an agar gel immunodiffusion (AGID) assay for detection of myxoma virus (MYXV) in the classical form of myxomatosis and to compare its diagnostic performance to that of molecular methods (IAC-PCR, OIE PCR, and OIE real-time PCR). A panel of MYXV-positive samples of tissue homogenates with low (1 PCR unit – PCRU) and high (3,125 PCRU) virus levels and outbreak samples were used for method comparison studies. The validation parameters of the AGID assay were assessed using statistical methods. The AGID attained DSe of 0.65 (CI₉₅%: 0.53–0.76), DSp of 1.00 (CI₉₅%: 0.40–1.00), and accuracy of 0.67 (CI₉₅%: 0.55–0.76). The assay confirmed its diagnostic usefulness primarily for testing samples containing ≥3,125 PCRU of MYXV DNA. However, in the assaying of samples containing <3,125 PCRU of the virus there was a higher probability of getting false negative results, and only molecular methods showed a 100% sensitivity for samples with low (1 PCRU) virus concentration. The overall concordance of the results between AGID and IAC-PCR was fair (ĸ = 0.40). Full concordance of the results was observed for OIE PCR and OIE real-time PCR when control reference material was analysed. Findings from this study suggest that AGID can be used with some limitations as a screening tool for detection of MYXV infections.

Early detection of pregnancy is vital for appropriate reproductive management programmes to facilitate the rapid re-insemination of non-pregnant females and reduce the calving interval. A barium chloride test was compared with a commercial progesterone ELISA to detect pregnancy in non-descriptive cows and investigate if it could be applied as an alternative to ELISA in the field. Blood and urine samples were collected from 74 cows with recorded insemination dates. The progesterone ELISA and barium chloride assay were implemented to detect progesterone (P4) in blood and urine specimens, respectively. The cows' reproductive systems were examined after they were slaughtered to determine the uterus's status. Macroscopic examination of the uterus was used as a reference standard for both tests. The sensitivity rates of the P4 ELISA and barium chloride test to detect pregnant cows were 100.0% and 79.4%, and to detect the corpus luteum (CL) were 83.0% and 87.0%, respectively, their sensitivity increasing in the presence of the CL. The ELISA and barium chloride tests were 79.7% and 52.7% accurate in the diagnosis of pregnancy. The accuracy of the barium chloride test in CL detection increased to 81.0%, and that of the ELISA to 86.4%. There were no significant differences (P = 0.052) between the barium chloride assay and ELISA when they were utilised for the identification of the CL. The barium chloride test can be an inexpensive and time-saving alternative to ELISA in pregnancy diagnosis when the insemination date is known.

Gastrointestinal parasites are some of the most common pathogens which are seriously harmful to the camel’s health. The infection status of gastrointestinal parasites in camels (Camelus bactrianus) in the Tianshan Mountains pastoral area in China is still unclear. The aim of this study was to investigate the species and infection intensity of gastrointestinal tract parasites in local camels. A total of 362 fresh faecal samples were collected and examined for parasite eggs using the saturated saline floating and natural sedimentation method. The parasite eggs were subjected to morphological and molecular examination and identification, and the infection rate and mean intensity of the parasites were analysed. A total of 15 gastrointestinal tract parasite species’ eggs were identified, with a detection rate of 100%. Ostertagia spp. (100%) and Trichostrongylus spp. (98.1%) were dominant. Camels were often coinfected by 5–14 species. The average number of eggs per gram of faeces was higher for Ostertagia spp. (298), Haemonchus contortus (176) and Nematodirus spp. (138). The number of species of parasites infecting young camels was significantly lower than that of adult camels, but the infection intensity in young camels was significantly higher. Gastrointestinal parasites were highly prevalent in camels from the Tianshan Mountains pastoral area in China. This finding provides important epidemiological data for the prevention and control of associated infections in camels.

Tetrodotoxin (TTX) is a toxin mainly occurring naturally in contaminated puffer fish, which are a culinary delicacy in Japan. It is also detected in various marine organisms like globefish, starfish, sunfish, stars, frogs, crabs, snails, Australian blue-ringed octopuses, and bivalve molluscs. TTX is produced by marine bacteria that are consumed mainly by fish of the Tetraodontidae family and other aquatic animals. TTX poisoning through consuming marine snails has recently begun to occur over a wider geographical extent through Taiwan, China, and Europe. This neurotoxin causes food intoxication and poses an acute risk to public health. The aim of this review is to present the most recent information about TTX and its analogues with particular regard to toxicity, methods of analysis, and risk to humans of exposure.