Circulation dynamics of Salmonella enterica subsp. enterica ser. Gallinarum biovar Gallinarum in a poultry farm infested by Dermanyssus gallinae.

Dermanyssus gallinae (Mesostigmata: Dermanyssidae, De Geer, 1778) is an ectoparasite of poultry, suspected to play a role as a vector of Salmonella enterica subsp. enterica ser. Gallinarum. Despite an association between them being reported, the actual dynamics in field remain unclear. Therefore, the present study aimed to confirm the interactions among mites, pathogen and chickens. The study was carried out in an industrial poultry farm infested by D. gallinae, during an outbreak of fowl typhoid. The presence of S. Gallinarum in mites was assessed and quantified by a semi-nested polymerase chain reaction (PCR) and real-time PCR, respectively, in mites collected during two subsequent productive cycles and the sanitary break. The anti-group D Salmonella antibodies were quantified by an enzyme-linked immunosorbent assay. During the outbreak and the sanitary break, S. Gallinarum was constantly present in mites. In the second cycle, scattered positivity was observed, although hens did not exhibit signs of fowl typhoid, as a result of the vaccination with BIO-VAC SGP695 (Fatro, Ozzano Emilia Bo, Italy). The data strongly suggest that D. gallinae acts as reservoir of S. Gallinarum, thus allowing the pathogen to persist in farms. Furthermore, the present study has highlighted the interactions among D. gallinae, S. Gallinarum and hens with respect to enhancing the mite-mediated circulation of S. Gallinarum in an infested poultry farm.

Chlamydia psittaci infection in canaries heavily infested by Dermanyssus gallinae.

Dermanyssus gallinae is a haematophagous ectoparasite responsible for anemia, weight loss, dermatitis and a decrease in egg production. Dermanyssus gallinae may play a role in the modulation of the host immune system, maybe predisposing the host to some bacterial infections such as chlamydiosis. This is an important zoonosis. Humans are exposed to Chlamydia psittaci through inhalation of the agent dispersed from the infected birds. In this study, a syndrome observed in an aviary of canaries was investigated. A heavy infestation by D. gallinae was reported. Simultaneously, a C. psittaci infection was molecularly confirmed in the canaries. Combined therapy was applied successfully. The association of C. psittaci with the examined mites has been confirmed. Therefore, we think that D. gallinae have played a role in the spreading of C. psittaci infection among the canaries. Moreover, D. gallinae could have played an important role predisposing the canaries to the development of chlamydiosis, by inducing anemia and debilitation. The control of mites in the aviaries may represent a crucial step for the prevention of important infection such as chlamydiosis in birds and humans.

Characterization of Dermanyssus gallinae (Acarina: Dermanissydae) by sequence analysis of the ribosomal internal transcribed spacer regions.

In the present work mites previously identified as Dermanyssus gallinae De Geer (Acari, Mesostigmata) using morphological keys were investigated by molecular tools. The complete internal transcribed spacer 1 (ITS1), 5.8S ribosomal DNA, and ITS2 region of the ribosomal DNA from mites were amplified and sequenced to examine the level of sequence variations and to explore the feasibility of using this region in the identification of this mite. Conserved primers located at the 3'end of 18S and at the 5'start of 28S rRNA genes were used first, and amplified fragments were sequenced. Sequence analyses showed no variation in 5.8S and ITS2 region while slight intraspecific variations involving substitutions as well as deletions concentrated in the ITS1 region. Based on the sequence analyses a nested PCR of the ITS2 region followed by RFLP analyses has been set up in the attempt to provide a rapid molecular diagnostic tool of D. gallinae.

Phylogenetic relationship between Dermanyssus gallinae populations in European countries based on mitochondrial COI gene sequences.

Phylogenetic analysis of Dermanyssus gallinae mites originating from UK, France and Italy was performed using partial amplification of the mitochondrial COI gene. Results showed that UK samples reveal the greatest variation and diversity and are linked to one of the French populations highlighting North-South genetic transitions in European red mite populations. Intra-farm variations between mite samples highlighted the diversity between national populations and possibly its origin from the different chemical strategies used in each country.

Evaluation of the poultry red mite, Dermanyssus gallinae (Acari: Dermanyssidae) susceptibility to some acaricides in field populations from Italy.

Red mite field populations from seven naturally infested Italian caged laying poultry farms were investigated for their susceptibility to acaricide formulations available on the market, containing amitraz, carbaryl and permethrin. A minimum of 3,000 mites of all stages were collected from each farm and were tested with five acaricide concentrations (5, 10, 20, 50, 100%) plus an untreated control (0%). Field red mite populations were found to be tolerant even with the highest concentrations with carbaryl and permethrin for six (86%) and three (42%) of the investigated farms, respectively (P < 0.05). Furthermore, six (86%) of the investigated farms showed a red mite population susceptible to amitraz at any concentration. Out of the seven field populations tested with amitraz, one population is becoming less tolerant whereas another was the most tolerant to carbaryl and permethrin at any concentration. Data show that the lack of effectiveness of some acaricides is spreading in Europe and call for the adoption of alternative management strategies to avoid development of resistance.

Bluetongue virus isolations from midges belonging to the Obsoletus complex (Culicoides, Diptera: Ceratopogonidae) in Italy.

Between July and September 2002 there were outbreaks of bluetongue on three sheep holdings in the communities of San Gregorio Magno (Salerno, Campania), Laviano (Salerno, Campania) and Carpino (Foggia, Puglia), and the involvement of bluetongue virus (btv) was confirmed serologically and virologically. The mortality rate was at least 11 per cent and involved btv serotype 2 (btv-2) and serotype 9 (btv-9). These holdings were also surveyed for the Culicoides (Diptera: Ceratopogonidae) vectors; approximately 10,000 midges belonging to 15 species were captured, but they did not include a single specimen of the classical Afro-Asiatic bluetongue vector, Culicoides imicola. Species belonging to the Obsoletus complex dominated the light-trap collections, and Culicoides obsoletus Meigen, Culicoides scoticus Downes and Kettle and Culicoides dewulfi Goetghebuer constituted 90 per cent of all the Culicoides species captured. Fifty-six pools of the Obsoletus complex (excluding C dewulfi), each containing 100 individual midges and containing only parous and gravid females, were assayed for virus. btv-2 was isolated from three pools from San Gregorio Magno and Carpino, and btv-9 was isolated from one pool from Laviano. These results indicate that a species other than C imicola is involved in the current re-emergence of bluetongue in the Mediterranean Basin, but whether it is C obsoletus sensu stricto or C scoticus, or both, is uncertain.

[Coxiella burnetii: what is the reality?]. / Coxiella burnetii: quale realtà?

After the Second World War, in Italy Q Fever or Coxiellosis has been shown a significant relevance, a recrudescence with an epidemic state for over ten years. Later, the infectious disease occurred as endemic since the 80s, the outbreaks were just isolated. Workflows analysis of some authors has demonstrated the spread out of the infection throughout Italian herds with a prevalence ranging from 1.2 per cent to 10 per cent. Our survey carried out throughout Campania area in cattle has shown a real positivity over 14 per cent performing the IFAT for the detection of IgG antibodies for Coxiella burnetii. Therefore, it has been so important to stress the influence of cattle farming management in stables as a real risk of Coxiellosis. For example, the Relative Risk (RR) has been registrated about 6.84 (2.18

Health management of large transhumant animal populations and risk of bluetongue spread to disease-free areas.

Transhumance, or seasonal grazing, in central Italy is a husbandry practice that is over two thousand years old. It involves the seasonal movement of sheep, goats and cattle from the southern lowlands of mainly the Puglia and Lazio regions, to summer pastures in the mountains of Abruzzo and Molise. Bluetongue (BT) made its appearance in Italy in 2000. In the early summer of 2001, disease was present in three regions: Sardinia, Sicily and Calabria. Neither an effective surveillance system nor a vaccination campaign had been implemented. Movement of ruminants to the disease-free regions of Abruzzo and Molise was therefore banned. The Italian Veterinary Services had to meet the challenge of the movement of ruminants from surveillance to disease-free zones, given the impossibility of stopping transhumance. The General Directorate of Veterinary Public Health, Food and Nutrition of the Ministry of Health developed a plan for both the Puglia and Abruzzo regions based on serological, virological and entomological surveillance. The plan was implemented between May and June 2001 when 7,000 animals moved from the Puglia surveillance zone to the infection-free summer pastures. In the early summer of 2002, eight regions were infected (Sardinia, Sicily, Calabria, Basilicata, Puglia, Campania, Lazio and Tuscany). Simultaneously, a nationwide surveillance system and a vaccination campaign, were implemented in infected regions. In the provinces where vaccination was compulsory, deviation from the animal movement ban was allowed if at least 80% of susceptible stock had been vaccinated. However, this objective was not achieved in the provinces of Rome and Viterbo (Lazio) where a large transhumant population was present and where sporadic virus circulation had been detected. A specific control plan to allow transhumance from Lazio to Abruzzo, Marche and Umbria was designed and implemented to increase the number of animals that could be moved. Between May and June 2002, authorisation was granted to move 28,000 head, whereas prohibition of movement was ordered for 12,000 sheep (belonging to 21 flocks). Regional authorities financed feeding, watering and housing for these animals. Transhumance did not spread infection to disease-free areas either in 2001 or in 2002.

Molecular analysis of the VP7, VP4, VP6, NSP4, and NSP5/6 genes of a buffalo rotavirus strain: identification of the rare P[3] rhesus rotavirus-like VP4 gene allele.

We report the detection and molecular characterization of a rotavirus strain, 10733, isolated from the feces of a buffalo calf affected with diarrhea in Italy. Strain 10733 was classified as a P[3] rotavirus, as the VP8* trypsin cleavage product of the VP4 protein revealed a high amino acid identity (96.2%) with that of rhesus rotavirus strain RRV (P5B[3]), used as the recipient virus in the human-simian reassortant vaccine. Analysis of the VP7 gene product revealed that strain 10733 possessed G6 serotype specificity, a type common in ruminants, with an amino acid identity to G6 rotavirus strains ranging from 88 to 98%, to Venezuelan bovine strain BRV033, and Hungarian human strain Hun4. Phylogenetic analysis based on the VP7 gene of G6 rotaviruses identified at least four lineages and an apparent linkage between each lineage and the VP4 specificity, suggesting the occurrence of repeated interspecies transmissions and genetic reassortment events between ruminant and human rotaviruses. Moreover, strain 10733 displayed a bovine-like NSP4 and NSP5/6 and a subgroup I VP6 specificity, as well as a long electropherotype pattern. The detection of the rare P[3] genotype in ruminants provides additional evidence for the wide genetic and antigenic diversity of group A rotaviruses.