Beyond Borders: Dirofilaria immitis Infection in Dogs Spreads to Previously Non-Enzootic Areas in Greece-A Serological Survey.

Although Dirofilaria immitis in dogs is considered enzootic in northern Greece, the available data on the occurrence of infection in southern parts of the country demonstrate its scarcity. The aim of this study was to update the current knowledge on D. immitis infection in dogs in areas of Greece previously considered non-enzootic (Central Greece, Attica, Peloponnese, North Aegean, South Aegean, Crete and the Ionian islands). In total, 1528 dog blood samples were collected from the aforementioned areas and examined by Dirochek® ELISA (Synbiotics). Additionally, data published until 2022 on the prevalence of infection in these areas were compared to the data of this study. The 'Wilson' Score interval method (Epitools) was employed. Overall, 10.8% of dogs were positive for D. immitis. In detail, the prevalence was 21.7, 13.7, 10.7, 5.4, 4.7, 6.2 and 17.0% for D. immitis, in Central Greece, Attica, Peloponnese, North Aegean, South Aegean, Crete and the Ionian islands, respectively. Infection with D. immitis is recorded for the first time in Crete. The probability of a dog becoming infected has increased 4.1 times since 2022 in previously non-enzootic areas. This study denotes the spread of D. immitis and highlights the necessity for preventive measures.

Is the European badger a new host for Dirofilaria immitis? The first records in Greece.

Dirofilaria immitis is a ubiquitous nematode parasite with zoonotic potential, transmitted by mosquitoes, that causes heartworm disease in various animal species. Dogs are the parasite's typical final host, and wild carnivores represent the parasite's reservoir in nature. Studies on D. immitis infections in wild animals are essential to assess infection pressure for domestic animals, and until now, there has been only one infection case reported in a European badger (Meles meles). The current report describes the first two European badger cases with cardiovascular dirofilariosis in Greece. Two adult male badgers were rescued in Heraklion and Chania, Crete Island, and admitted to "ANIMA -Wildlife Rehabilitation Centre" in Athens. The detailed clinical examination revealed that the first badger suffered from severe broncho-pneumonitis while the second one displayed clinical signs associated with severe brain trauma. Blood samples were taken for haematology and biochemistry analyses during their short hospitalisation period. In addition, different routine diagnostic tests were carried out, including heartworm antigen testing (ELISA) and the modified Knott's test for microfilariae. Both badgers were positive in both tests. The animals died a few hours after their admission and the detailed necropsies followed, revealed the presence of three parasites in each animal's right heart, morphologically identified as adults of D. immitis. These findings add the European badger in the list of additional potential reservoir hosts for D. immitis and highlight the potential role of wildlife for companion animals and human health.

Evaluation of porcine circovirus type 2 double vaccination in weaning piglets that reared for gilts under field conditions.

The objective of the present study was to evaluate the efficacy of a porcine circovirus type 2 (PCV2) double vaccination in weaning piglets reared for gilts under field conditions. The study was conducted at a Greek farrow-to-finish conventional pig farm with a previous history of PCV2 infections. The trial included 96 female piglets at 21 days of age, which were equally allocated to two different study groups. Piglets of the group-1 received a single PCV2 vaccination at 21 days of age, while piglets of the group-2 were double vaccinated against PCV2, at 21 and 42 days of age. The results indicated that the piglets of group-2 had better growth performance, as they showed higher body weight (BW) and average daily weight gain (ADWG). In addition, ELISA tests showed that the double- vaccinated piglets presented a better humoral response against PCV2, as higher levels of IgG antibodies were detected in them than the piglets of the group-1. In conclusion, the current results suggested that a double PCV2 vaccination of piglets, reared for gilts, on a PCV2-affected farm could lead to higher protection against the virus.

Occurrence and Associated Risk Factors of Porcine Reproductive and Respiratory Syndrome Virus and Porcine Circovirus Type 2 Infections in Greece.

The objective of the present study was to identify factors associated with the probability of being polymerase chain reaction (PCR) positive and the level of porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2) occurrence in Greek farms. The study included 56 pig farms, with a total population of 22.500 sows, which represent about 40% of the entire capacity of the Greek swine production. A total of 896 blood samples (224 pools*4 samples/pool = 896 samples) from breeding stock, weaners, growers, and finishers were collected from each farm and organized in pools of 4 samples size. Further, data regarding herd health management protocols were collected. The sera were tested for PRRSV and PCV2, using real-time PCR (RT-PCR). The results indicated that both viruses remain a major challenge for the Greek swine industry. Main risk factors involved in the infection process by these viruses were identified. In particular, vaccination programs such as the mass PRRSV vaccination with modified-live virus (MLV) in breeding stock during the last stages of gestation or with killed-virus (KV) during the middle of gestation are more likely to be associated with PRRSV PCR-positivity. Farms with low biosecurity level are associated with higher PRRSV circulation. It has also been revealed that breeding stock is more likely to be associated with PCV2 circulation compared to weaners and growers. In conclusion, our results could be the basis of the development of surveillance protocols for a national monitoring system for PRRSV and PCV2, which could prevent future infection of Greek farms.

Effect of Toxoplasma gondii on Ram Sperm Quality After Experimental Infection.

The aim of this study was to investigate the effect of experimental Toxoplasma gondii infection on ram sperm quality. Five months old, pre-pubertal, rams were divided into four groups (n = 8 per group). Group A was the control group; the remaining animals received per os (p.o.) 5000 oocysts per ram. Group B did not receive treatment post-infection (p.i.). Group C received sulphadimidine (intermuscular injection (i.m.) 33 mg/kg for eight days; every 48 hrs) two months p.i. and Group D received the same drug twice (24 hours p.i. and two months later). Blood samples were collected every 15 days to detect serum immunoglobulin G (IgG). Epididymal sperm samples were analyzed for concentration, kinetics, morphology/viability, functional membrane integrity, DNA integrity, and the presence of parasite DNA. Histopathological examination was performed on the testes. The IgG titres in infected groups raised two weeks p.i. and remained high for four months. Higher values were noticed in viability and functional membrane integrity in positive spermatozoa in the control group compared to other groups, level of significance p < 0.05. Abnormal sperm was higher in groups C and D vs. A and C vs. B (p < 0.05). T. gondii DNA was detected in three sperm samples of the infected rams (12.5%). Histopathology revealed similar findings with little variation among all infected groups, characterized mostly by increased interstitial connective tissue, non-purulent inflammation, and presence of seminiferous tubules with spermatogenic cell depletion, which increased gradually from D to C and B groups. In conclusion Toxoplasmosis in pre-pubertal age negatively affected mature ram sperm quality, while sulphadimidine administration failed to alter this.

Histopathological Lesions Accompanied with First-Time Isolation of a PRRSV-2 Strain in Greece.

Genotype 2 strains of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV-2) have been reported sporadically in Europe. Even if, PRRSV-2 reported to be genetically homogenous in Europe due to the introduction of an MLV vaccine strain, independent introductions of PRRSV-2 field strains have been reported. The aim of the present study was to report the complete genome sequence and evaluate the histopathological lesions of a PRRSV-2 strain, isolated for the first time in Greece. During a routine blood sampling in a commercial pig farm, the results revealed positive samples in weaners of 40-60 days for the PRRSV-2, using real-time polymerase chain reaction. The clinical picture was characterized from respiratory symptoms in weaners, as well as coughing and poor performance at finishing stage and less than 3% mortality rate from weaning stage to finishing stage. The use of ORF5 for PRRSV phylogenetic analysis of the isolated PRRSV strain, named "x1544-1 strain", was successfully determined, belonging to the genotype PRRSV-2. Comparison of the obtained sequence revealed nucleotide sequence identity >98% with PRRSV-2 strain VR2332 and other related strains from Denmark and China. The histopathological evaluation revealed diffuse interstitial pneumonia, multifocal interstitial nephritis, while in the lymphoid organs, follicular and paracortical hyperplasia, coexisting with necrosis and depletion of germ cells were detected. The results of current study undersign the importance for veterinary practitioners to have up-to-date access to phylogenetic data linked to phenotypic information to follow-up the control and prevention strategies against PRRSV.