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Background: Encephalitozoon cuniculi can affect a number of animals. Currently, there is limited clinical information on E. cuniculi in guinea pigs. Case description: A 3-year-old guinea pig with torticollis, dehydration, and hypothermia was brought to the clinic. After symptomatic treatment, the animal's status improved, and the animal returned home. Two computed tomography scans over a period of 23 days revealed oval hypodense lesions in the right temporal lobe, which enlarged over time. The animal was euthanized. A histopathological examination revealed focal granulomatous encephalitis, and PCR allowed the detection of E. cuniculi DNA in the brain tissue. It was found that the possible cause of the disease was E. cuniculi invasion. Conclusions: Based on this report, E. cuniculi should be considered a differential in guinea pigs with granulomatous encephalitis.

Introduction The prohibition of antibiotic use in edible snails obligates breeders to treat bacterial infections by different means, of which a common one is a bath in Gram-positive– and partially Gram-negative–bactericidal ethacridine lactate solution. The aim of the study was to determine the effect of bathing Cornu aspersum Müller snails in a 0.1% aqueous solution of ethacridine lactate on selected physiological parameters of haemolymph. Material and Methods The study included 80 snails, divided into two equal groups (study and control). The study group was subjected to bathing in ethacridine lactate and the control group to bathing in tap water. Both groups were treated daily for seven days. The number of haemocytes in the haemolymph, the activity of alanine (ALT) and aspartate (AST) aminotransferases, and the concentration of urea were determined. Results In the study group, after exposure to ethacridine lactate solution an increase in ALT activity, changes in the De Ritis ratio, an increase in the amount of haemocytes, and a decrease in body weight were found. No such changes were detected in the control group snails or in animals after the first bath. Conclusion Multiple applications of a 0.1% ethacridine lactate bath may adversely affect Cornu aspersum Müller snails.

Canine parvovirus type 2 is one of the most common causes of death among puppies. Despite preventive vaccination, the disease continues to be diagnosed. The aim of the study was to provide a molecular characterization of CPV-2 isolates found in southeastern Poland. Genetic CPV-2 material was isolated from the blood (n=10) and feces (n=50) of infected dogs. The presence of CPV-2 was confirmed by amplification of sequences coding both VP1 and VP2 protein. The products of the PCR reaction with primers amplifying VP2 protein were sequenced and used for genotyping. Bioinformatics analysis of the sequenced PCR product was performed to determine the phylogenetic relationships with variants recorded in the public databases. Based on the analysis of polymorphism in the nucleotide sequence 7 nucleotide variants were detected and assigned into four amino acid groups. Representatives of three groups contained asparagine at amino acid position 426 of the VP2 protein, which is characteristic of CPV-2a. The variant from the fourth group belonged to type CPV-2b. CPV-2a is the dominant antigenic type of CPV-2 in Poland. The pathogen’s high degree of polymorphism is manifested not only by the presence of numerous variants within the type, but also by the presence of representatives of type CPV-2b. Further studies of the molecular epidemiology of CPV-2 are necessary to optimize the effectiveness of preventive measures.

Background Anaplasma are obligate intracellular bacteria and aetiological agents of tick-borne diseases of both veterinary and medical interest. The genus Anaplasma comprises six species: Anaplasma marginale, Anaplasma centrale, Anaplasma ovis, Anaplasma phagocytophilum, Anaplasma bovis and Anaplasma platys. They can infect humans, carnivores, ruminants, rodents, insectivores, birds and reptiles. The aim of this study was to present the first clinical case of granulocytic anaplasmosis in a captive ring-tailed lemur in Poland. Case presentation A 4-year-old female lemur presented anorexia, epistaxis and tick infestation. The microscopic examination of a blood smear revealed morulae in neutrophils. Polymerase chain reaction test and sequencing of obtained PCR product confirmed infection by the GU183908 Anaplasma phagocytophilum strain. Therapeutic protocol included doxycycline (2.5 mg/kg p.o., b.i.d.) for 3 weeks and the lemur recovered within 24 h. Conclusions This is the first report on granulocytic anaplasmosis in a ring-tailed lemur in Europe, indicating that A. phagocytophilum infection must also be considered in differential diagnosis in this animal species, especially in individuals with thrombocytopenia associated with Ixodes ricinus parasitism.

Chitin, as one of nature&rsquo, s most abundant structural polysaccharides, possesses worldwide, high industrial potential and a functionality that is topically pertinent. Nowadays, the metallization of naturally predesigned, 3D chitinous scaffolds originating from marine sponges is drawing focused attention. These invertebrates represent a unique, renewable source of specialized chitin due to their ability to grow under marine farming conditions. In this study, the development of composite material in the form of 3D chitin-based skeletal scaffolds covered with silver nanoparticles (AgNPs) and Ag-bromide is described for the first time. Additionally, the antibacterial properties of the obtained materials and their possible applications as a water filtration system are also investigated.

Structure-based tissue engineering requires large-scale 3D cell/tissue manufacture technologies, to produce biologically active scaffolds. Special attention is currently paid to naturally pre-designed scaffolds found in skeletons of marine sponges, which represent a renewable resource of biomaterials. Here, an innovative approach to the production of mineralized scaffolds of natural origin is proposed. For the first time, a method to obtain calcium carbonate deposition ex vivo, using living mollusks hemolymph and a marine-sponge-derived template, is specifically described. For this purpose, the marine sponge Aplysin aarcheri and the terrestrial snail Cornu aspersum were selected as appropriate 3D chitinous scaffold and as hemolymph donor, respectively. The formation of calcium-based phase on the surface of chitinous matrix after its immersion into hemolymph was confirmed by Alizarin Red staining. A direct role of mollusks hemocytes is proposed in the creation of fine-tuned microenvironment necessary for calcification ex vivo. The X-ray diffraction pattern of the sample showed a high CaCO3 amorphous content. Raman spectroscopy evidenced also a crystalline component, with spectra corresponding to biogenic calcite. This study resulted in the development of a new biomimetic product based on ex vivo synthetized ACC and calcite tightly bound to the surface of 3D sponge chitin structure. © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). Naturalis Biodiversity Center Ministerio de Ciencia, Tecnología y Medio Ambiente, CITMA: 18-54-34007 Deutsche Forschungsgemeinschaft, DFG: HE 394/3 Russian Foundation for Basic Research, RFBR Russian Science Foundation, RSF: 18-13-00220 57397326 Ministerstwo Nauki i Szkolnictwa Wyższego, MNiSW: 03/32/SBAD/0906 PPN/BEK/2018/1/00071 Deutscher Akademischer Austauschdienst, DAAD Sächsisches Staatsministerium für Wissenschaft und Kunst, SMWK: 02010311 Funding: This work was partially supported by the DFG Project HE 394/3, SMWK Project no. 02010311 (Germany), as well as DAAD-Italy Project “Marine Sponges as Sources for Bioinspired Materials Science” (No. 57397326). The sampling conducted in Cuba (V.I.) and Curaçao (V.I. and B.H.) was funded by RFBR and CITMA, according to the research project 18-54-34007 (grant), and Temminck-Fellowships from the Naturalis Biodiversity Center, respectively. M.W is financially supported by the Polish National Agency for Academic Exchange (PPN/BEK/2018/1/00071). Tomasz Machałowski is supported by DAAD (Personal ref. no. 91734605). Yuliya Khrunyk is supported by the Russian Science Foundation (grant no. 18-13-00220). This research was partially funded by the Ministry of Science and Higher Education (Poland), as financial subsidy to PUT No. 03/32/SBAD/0906.

The aim of the study was to develop an intravital method of collecting hemolymph and to establish standards for the biochemical parameters of the hemolymph of the Cornu aspersum snail species. The research material consisted of 80 Cornu aspersum maxima snails. Hemolymph was collected from the main vessel with a sterile syringe and needle after removing a fragment of the shell. Hemolymph was studied biochemically by the colorimetric method. The parameters evaluated were aspartate transaminase (AST), alanine transaminase (ALT), amylase (AMYL), urea (UREA), and triglycerides (TG). An average of 0.54 ml hemolymph was collected from the animals tested. After 24 hours, a film formed in place of the removed shell fragment and it underwent mineralization within a few days. All animals included in the study survived, and from some of them it was possible to collect hemolymph several times at intervals of several days. The proposed method of hemolymph collection made it possible to obtain material to develop standards for the biochemical parameters of hemolymph and consequently to monitor the health status of these mollusks.

The aim of this study was to use matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of coagulase-negative staphylococci (CNS) isolated from the milk of cows with subclinical mastitis. The study material consisted of 33 isolates of CNS, identified by the results of API Staph tests, obtained from the milk of cows with subclinical mastitis. Based on the spectra analyses, MALDI-TOF MS tests of 33 bacterial samples allowed identification of the microorganisms in 27 cases (81.8%). The most frequent cause of subclinical mastitis was found to beStaphylococcussciuri (39%), whileS. vitulinuswas detected in 15% of the milk samples. The results obtained indicate that MALDI-TOF MS can be used for the identification of CNS isolated from bovine mastitis as a method supplementary to biochemical tests.

Snails of the genus Cornu are farm-raised as edible molluscs. Dissection is one of the basic diagnostic tests available in the breeding of these animals, used to determine the cause of death or disease, and to collect material for further laboratory tests. The aim of this article was to present a method for the dissection of snails for veterinary use based on the experience gained from 200 mollusc dissections, and to present a short description of the anatomical structure of snails. The method described is characterized by its speed and simplicity. The observations made during the dissection, as presented in the article provide valuable diagnostic information for veterinarians responsible for care on snail farms.

Endophytes are fungi of the genus Neotyphodium that live in symbiosis with grasses. The effects of this symbiosis can be both negative and positive. The negative effect may be due to alkaloids produced by endophytes that in higher concentrations are toxic to animals. The most common alkaloids produced by Neotyphodium fungi are ergovaline and lolitrem B. The current state of knowledge on the settlement of grasses by these fungi in Poland is rather small. It has been observed that fungi from the genus Neotyphodium occurring in a pasture sward can produce alkaloids. The average ergovaline content in grass from the permanent pasture in some areas exceeded the threshold level above which disease symptoms in animals may occur. The aim of this paper was to present a clinical case of ergovaline intoxication in a herd of 30 dairy cows. Out of the 30 animals, in 10 self amputation of the tails and visual disturbances were observed. Ergovaline poisoning was diagnosed on the basis of the results of analysis of the sera of infected animals by mass spectrometer. It enabled detecting a specific fraction in the serum of sick cows that was identified as ergovaline. The case of ergovaline poisoning confirmed by laboratory tests indicates that in some areas poisoning with this alkaloid may be a significant problem in the breeding of ruminant animals, which should be considered in the differential diagnosis of diseases with necrosis of the terminal parts of the body and decreased milk production

The purpose of the study was the in vivo diagnosing of E. cuniculi invasions in pet rabbits with neurological symptoms using the Real-Time PCR, and determination of the rate of invasion, in this group of animals. The study involved 103 pet rabbits with neurological symptoms. Parasitic invasions were diagnosed using Real-Time PCR. The DNA of the parasites for molecular tests was isolated from the urine of the diseased animals. Out of the 103 tested DNA samples, the presence of the E. cuniculi genetic material was detected in 27 samples (26.21%). The melting temperature (Tm) of all products was 77.5oC. The presence of parasitic DNA in the urine of 26.21% of examined animals indicates that E. cuniculi infections occur widely in pet rabbits in Poland and are a significant cause of neurological disorders in those animals

The aim of the present study was to investigate the occurrence of Anaplasma spp. in group of 50 fallow deer (Dama dama) from free-range farm in eastern Poland and determine what species of Anaplasma could infect these animals based on PCR gene sequencing. The PCR technique revealed the presence of 16S RNA Anaplasma spp. genetic material in the blood of 7 out of 50 examined animals. The sequences of the PCR products obtained showed a 100% homology with each other and 100% homology with GU 183908 sequence of A. phagocytophilum, isolated in our earlier study from a horse with clinical form of anaplasmosis. Here, we report the first molecular evidence of Anaplasma spp. among naturally infected fallow deer in eastern Poland.

Canine parvovirus disease appeared in the world and in Europe during the second half of the 1970s. Over the course of 40 years the original CPV-2 strains mutated and variants 2a, 2b and 2c appeared. Their appearance is connected with specific amino acid changes, mainly in the capsid protein VP2. Strains isolated by the authors were adapted for in vitro cell culture. Phylogenetic analysis revealed differences between strains isolated in Poland in 1982-1985 and in 1995-2009. Strains from the 1980s were shown to belong to variant CPV-2a (11 strains) and variant 2b (2 strains), while no fundamental differences were found among the genetic profiles of the strains from 1995-2009, which were classified as belonging to variant 2c.