Psychrobacter raelei sp. nov., isolated from a dog with peritonitis.

A strain belonging to the genus Psychrobacter, named PraFG1T, was isolated from the peritoneal effusion of a stray dog during necropsy procedures. The strain was characterized by the phylogenetic analyses based on the nucleotide sequences of 16S and 23S rRNA genes and of gyrB, which placed the strain in the genus Psychrobacter. The nucleotide sequence of the chromosome confirmed the placement, showing an average nucleotide identity of 72.1, 77.7, and 77.5 % with the closest related species, namely Psychrobacter sanguinis, Psychrobacter piechaudii, and Psychrobacter phenylpyruvicus, respectively, thus indicating a novel species. The polyphasic characterization by biochemical and fatty acid profiling as well as MALDI-TOF supported those findings. The strain was halotolerant, capable of growing within a temperature range between 4 and 37 °C, it was positive for catalase and oxidase, indole producing, nitrate reducing, and not able to use 5-keto-d-gluconic acid as a carbon source. Taken together, the data suggest that strain PraFG1T could be considered as representing a novel species, with the name Psychrobacter raelei sp. nov. (type strain PraFG1T=CIP 111873T=LMG 32233T).

Poisoning by Nerium oleander L. in Franconia Geese.

This study describes the acute poisoning of four 3-month-old Franconia geese (Anser anser) by oleander plants (Nerium oleander). After the accidental ingestion of oleander clippings, the geese exhibited a rapid onset of severe symptoms, leading to mortality within 15-90 min. Necropsy revealed cardiac and renal lesions. Specifically, interstitial edema, red blood cell infiltration, and myofibril loss were observed in the cardiac muscle, and tubular epithelial degeneration, interstitial edema, and hemorrhages were evident in the kidneys. Oleandrin, a glycoside with cardiac effects, was detected in the liver, kidneys, heart, brain, and muscles. The clinical implications underscore the urgency of veterinary intervention upon oleander ingestion, and the specific findings contribute valuable insights into the pathological effects of acute oleander poisoning in geese, aiding veterinarians in prompt diagnosis and treatment.

A Safe and Effective Atovaquone-Proguanil Therapeutic Protocol for the Treatment of Avian Malaria by Plasmodium relictum in Snowy Owl (Bubo scandiacus).

Avian malaria is a re-emerging threat to avian species worldwide. It is sustained by several protozoan species belonging to the genus Plasmodium, mainly Plasmodium relictum. The even wider diffusion of the disease, probably because of the increase in the areas covered by their mosquito vectors, may pose new risks for avian species lacking natural resistance (especially those from artic or sub-artic environments) or those hosted in structures like zoos and wildlife rescue centers. With that premise, this study describes the efficacy and safety of a therapeutic protocol to treat avian malaria in three snowy owls (Bubo scandiacus) hosted in a wildlife rescue center in Apulia, south of Italy, and affected by avian malaria by P. relictum. The protocol consisted of administering 10/4 mg/kg atovaquone/proguanil per os once a day for three consecutive days, repeating this seven days later. Seven days after the end of the treatment, P. relictum was not detected in the birds' blood and no adverse effects were observed during the 60 days of monitoring after the end of the treatment. Therefore, a therapeutic regimen of 10/4 mg/kg/day may be considered safe and effective in a valuable and endangered species such as B. scandiacus.

Profiling of Dermanyssus gallinae genes involved in acaricide resistance.

The poultry red mite (PRM), Dermanyssus gallinae, is a major threat for the poultry industry worldwide. Chemical compounds have been extensively used for PRM control, leading to selection of resistant mites. Molecular mechanisms of resistance have been investigated in arthropods, showing the role of target-site insensitivity and enhanced detoxification. Few studies are available about those mechanisms in D. gallinae, and none have yet focused on the expression levels of detoxification enzymes and other defense-related genes through RNA-seq. We tested PRM populations from Italy for their susceptibility to the acaricidal compounds phoxim and cypermethrin. Mutations in the voltage-gated sodium channel (vgsc) and in acetylcholinesterase (AChE) were investigated, detecting mutations known to be associated with acaricide/insecticide resistance in arthropods, including M827I and M918L/T in the vgsc and G119S in the AChE. RNA-seq analysis was performed to characterize metabolic resistance in fully susceptible PRM and in cypermethrin-resistant PRM exposed and unexposed to cypermethrin as well as phoxim resistant PRM exposed and unexposed to phoxim. Detoxification enzymes (including P450 monooxygenases and glutathione-S-transferases), ABC transporters and cuticular proteins were constitutively overexpressed in phoxim and cypermethrin resistant mites. In addition, heat shock proteins were found constitutively and inductively upregulated in phoxim resistant mites, while in cypermethrin resistant mites esterases and an aryl hydrocarbon receptor were constitutively highly expressed. The findings suggest that acaricide resistance in D. gallinae is due to both target-site insensitivity and overexpression of detoxification enzymes and other xenobiotic defense-related genes, which is mostly constitutive and not induced by treatment. Understanding the molecular basis of resistance could be useful to screen or test PRM populations in order to select targeted acaricides and to avoid the abuse/misuse of the few available compounds.

Pilot Study on the Action of Thymus vulgaris Essential Oil in Treating the Most Common Bacterial Contaminants and Salmonella enterica subsp. enterica Serovar Derby in Poultry Litter.

The indiscriminate use of antimicrobials in poultry farms is linked to the increase in multi-resistant bacteria. Accordingly, based on the antimicrobial properties of Thyme Essential Oil (TEO), the present study evaluated the effects of TEO on the reduction of common microbial contaminants and Salmonella on poultry litter. A litter bulk sample was collected in a broiler farm and qualitative/quantitative investigations identified Escherichia coli and Mammaliicoccus lentus. The experimental contamination with Salmonella Derby wild strain was also performed. All pathogens showed phenotypic and genotypic resistance to different classes of antibiotics. The litter, split in different units, was treated with aqueous solutions of TEO at different concentrations (5% to 1.25%), demonstrating its effectiveness in reducing the total number of bacteria. The strongest antibacterial action was observed at the lowest concentration against Enterobacteriaceae, with a growth reduction compared to the positive control of 73.3% and 77.8% against E. coli and Salmonella Derby, respectively, while towards M. lentus the reduction was 50%. Our data confirm the antimicrobial activity of TEO and suggest its possible application for the treatment of poultry litter as an effective and natural approach for the prevention of diseases caused by the most common bacteria that colonize poultry farms, counteracting the onset of antibiotic resistance.

Pseudomonas fluorescens group bacteria as responsible for chromatic alteration on rabbit carcasses. Possible hygienic implications.

Bacteria belonging to the genus Pseudomonas are ubiquitous and characterized by a high adaptation capability to different environmental conditions and wide range of temperatures. They may colonize food, sometimes causing alteration. Quite recently, a blue pigmentation due to Pseudomonas fluorescens has been widely reported in mozzarella cheese. In this report, we describe a blue coloration occurred on rabbit meat stored in the refrigeration cell of a slaughterhouse. The alteration was observed after about 72 hours of storage at 4-6°C. Bacteriological analyses were performed, and a microorganism included in the Pseudomonas fluorescens group was identified. The experimental contamination was planned, using a bacterial suspension with 1×108 UFC/ml load to spread on rabbit carcasses. The blue pigmentation appeared after 24 hours of storage in a cell with the same conditions of temperature. The bacterium was reisolated and identified as responsible for the alteration on meat. These findings highlight the importance of considering the members of the genus Pseudomonas and, more specifically, of the P. fluorescens group when the microbiological quality of food is to be ascertained. In fact, even if these bacteria are not considered a public health problem, their presence should be monitored by food industry operators in self-control plans because they may cause alteration in food. In fact, any altered product should be withdrawn from the market in agreement with Regulation (EC) No 178/2002 of the European Parliament and of the Council.

Release of DNA from Dermanyssus gallinae during the Biting Process.

Dermanyssus gallinae is a hematophagous ectoparasitic mite that usually infests poultry, but is also known for occasionally attacking other animals and humans. It represents a major problem for poultry systems all over the world, with detrimental effects for both production and animal welfare. Despite the significance of D. gallinae, very little is known about the biting process to date. Therefore, this study has aimed to verify if mite DNA is injected into the host skin during the blood meal. Mite DNA has been detected by seminested PCR from infested chicken skin and quantified by real-time PCR. Furthermore, its localization within the host tissue has been checked by fluorescent in situ hybridization. Results showed that a very little amount of D. gallinae DNA can be released by mites, suggesting that the latter do not introduce whole or partially destroyed cells into the host, but rather it injects traces of nucleic acids, possibly together with merocrine secretions.

Dermanyssus gallinae: the long journey of the poultry red mite to become a vector.

The possibility that Dermanyssus gallinae, the poultry red mite, could act as a vector of infectious disease-causing pathogens has always intrigued researchers and worried commercial chicken farmers, as has its ubiquitous distribution. For decades, studies have been carried out which suggest that there is an association between a wide range of pathogens and D. gallinae, with the transmission of some of these pathogens mediated by D. gallinae as vector. The latter include the avian pathogenic Escherichia coli (APEC), Salmonella enterica serovars Enteritidis and Gallinarum and influenza virus. Several approaches have been adopted to investigate the relationship between D. gallinae and pathogens. In this comprehensive review, we critically describe available strategies and methods currently available for conducting trials, as well as outcomes, analyzing their possible strengths and weaknesses, with the aim to provide researchers with useful tools for correctly approach the study of the vectorial role of D. gallinae.

Association between the poultry red mite Dermanyssus gallinae and potential avian pathogenic Escherichia coli (APEC).

Avian pathogenic Escherichia coli (APEC) is a group of strains responsible for avian colibacillosis, an impactful disease for poultry farming. The spread of APEC is mainly horizontal, and insects may play a role in their dissemination. However, no data are available about the interaction of APEC with Dermanyssus gallinae, a major arthropodal parasite of poultry. Escherichia coli was detected in the microbiome of the mite, but no specific data have been published till now. Therefore, the presence of the most diffused APEC-associated serogroups has been assessed by PCR in mites collected from 30 flocks of 21 Italian commercial laying hens farms. Escherichia coli was found in 53.3% of the tested groups, corresponding to 66.7% of farms. The most frequent serogroup was O2, but O8, O78, and O109 were also detected. More detailed investigations were carried out in a laying hen farm that was experiencing colibacillosis by APEC O2. The same serogroup was found in both hens and mites, and the maximum likelihood estimation (MLE) of the infection rate (IR) resulted in 24.39 infected mites per thousand, with a pathogen load of 171.47 E. coli O2 cells per mite. The results strongly support the hypothesis of an association between PRM and APEC, confirmed by the contemporary circulation of the same serogroup in both hens and mites, suggesting the potential of D. gallinae as a reservoir or APEC.

Pseudomonas azotoformans Belonging to Pseudomonas fluorescens Group as Causative Agent of Blue Coloration in Carcasses of Slaughterhouse Rabbits.

The study describes the finding of an abnormal blue-tinged color found on rabbit carcasses in the refrigeration cell of two butcher shops in Apulia Region. The carcasses were from an industrial rabbitry for production of meat with a regularly authorized slaughterhouse. Pseudomonas azotoformans, a microorganism included in Pseudomonas fluorescens group, was isolated from samples collected by the altered carcasses, showing the growth of uniform bacterial colonies with fluorescent pigmentation. The bacterium was also isolated from an additional water sample and from the labelling gun collected in the slaughterhouse, whilst the knives used for slaughtering resulted negative. Chromatic alteration was experimentally reproduced on new carcasses using a 108 cfu/mL bacterial suspension prepared with the isolated strain. Due to their resistance characteristics, members of P. fluorescens group are very difficult to eradicate once introduced into the production environment. Therefore, their presence, even if not considered a public health problem, should be monitored by food industry operators in self-control plans.