Co-Circulation of Different Hepatitis E Virus Genotype 3 Subtypes in Pigs and Wild Boar in North-East Germany, 2019.

Hepatitis E is a major cause of acute liver disease in humans worldwide. The infection is caused by hepatitis E virus (HEV) which is transmitted in Europe to humans primarily through zoonotic foodborne transmission from domestic pigs, wild boar, rabbits, and deer. HEV belongs to the family Hepeviridae, and possesses a positive-sense, single stranded RNA genome. This agent usually causes an acute self-limited infection in humans, but in people with low immunity, e.g., immunosuppressive therapy or underlying liver diseases, the infection can evolve to chronicity and is able to induce a variety of extrahepatic manifestations. Pig and wild boar have been identified as the primary animal reservoir in Europe, and consumption of raw and undercooked pork is known to pose a potential risk of foodborne HEV infection. In this study, we analysed pig and wild boar liver, faeces, and muscle samples collected in 2019 in Mecklenburg-Western Pomerania, north-east Germany. A total of 393 animals of both species were investigated using quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR), conventional nested RT-PCR and sequence analysis of amplification products. In 33 animals, HEV RNA was detected in liver and/or faeces. In one individual, viral RNA was detected in muscle tissue. Sequence analysis of a partial open reading frame 1 region demonstrated a broad variety of genotype 3 (HEV-3) subtypes. In conclusion, the study demonstrates a high, but varying prevalence of HEV RNA in swine populations in Mecklenburg-Western Pomerania. The associated risk of foodborne HEV infection needs the establishment of sustainable surveillance and treatment strategies at the interface between humans, animals, and the environment within a One Health framework.

[Post-mortem clinical pathology in cattle - an additional tool for the veterinary pathologist]. / Postmortale klinische Labordiagnostik beim Rind ­ ein Zusatzinstrument für die Veterinärpathologie.

Clinical-pathologic examination of samples collected from more or less preserved body compartments during necropsy may provide important information and contribute to the list of differential diagnoses without corresponding patho-morphologic findings. It furthermore allows for diagnoses that otherwise may only be achieved clinically. Our review presents diagnostic approaches in examining aqueous humor, urine, ruminal fluid as well as other sample types characterized by a delayed onset of auto- and heterolytic artefacts. An overview is provided concerning post mortem (p. m.) sample types and collection methods with a special focus on cattle. Furthermore, clinical-pathologic methods and parameters are presented and their validity discussed. A summary of pre-analytical caveats relevant for the final interpretation of findings is made available. Based on long-term experience in p. m. clinical pathology as well as literature information we provide practical approaches for daily routine diagnostics as well as for specific case scenarios. Especially aqueous humor, ruminal fluid, and urine are easily accessible sample types. The collection of high-quality cerebrospinal fluid allows for electrolyte and metabolite analyses. Post-mortem clinical pathology may provide an indication concerning the cause of death in specific cases e. g., recumbency and death due to hypocalcemia or hypomagnesemia. This is especially relevant in cases in which ante mortem clinical pathology investigations are hindered by rapid death of the animal as well as in cases in which the gross pathology findings cannot explain the clinical findings. Post-mortem clinical pathology may also be helpful when a clinical examination of the diseased animal is hampered. During necropsy for example uremia and ketosis may be detected based on their characteristic smell; however, the use of clinical chemical analyses allows the verification by an assessment of the metabolites in these cases. Post-mortem clinical pathology may hence help in establishing a diagnosis, narrow down the list of differentials or even reveal relevant differential diagnoses for the first time during the diagnostic process. Our review does not claim to be exhaustive; however, it serves to encourage the pathologist to make use of the so far rarely employed ancillary analyses as well as to promote the collaboration between veterinary and clinical pathologists.

Preanalytical errors on zinc concentrations caused by single-use gloves.

Preanalytical errors causing specimen contamination with zinc (Zn) are disastrous for routine medical diagnostics or scientific studies. The aim of the study presented here is to simulate contamination possibilities when using single-use gloves. The ability to release Zn into the specimen was tested using nitril (A), vinyl (B) and latex (C) gloves with 15 (14) replications. In our first approach, a 1 × 1 cm piece of the glove's fingertip was incubated for 10 min with serum. Our second approach imitated a very short contact of serum to the glove's material by letting serum run over the glove from a pipette for 3 cm distance into a tube. The effect of gloves' contact to liver tissue was examined using glove C only: a block of liver tissue was touched once at one side producing an experimental fingerprint. Zn was analyzed in serum and liver wet weight (ww) using ICP-MS; the basal serum/liver Zn concentration was set as zero for calculation. The calculated addition of Zn is given as median (p25 - p75). The first approach led to distinct contamination with Zn (in µg/L) being evident from all three types of gloves, but depended markedly from the type of material: A: 176.5 (129.7-204); B: 975.1 (663.6-1164.3); C: 2112 (1685-2516). Imitating a very short contact of serum to the glove's surface resulted in an additional Zn concentration of 105.7 (70.4-168.8), 56.2 (-13.5-121.4) and 955.7 (746-1159) µg/L using gloves A, B and C, respectively. A single fingerprint on liver tissue using glove C resulted an addition of 3995 (861-6435) µg Zn/kg liver ww. The data underline that the dimension of preanalytical contamination of blood and tissue samples for Zn analysis via single-use gloves is relevant for routine diagnostics and scientific studies. Critical steps and possibilities to minimize these effects should be considered seriously for specimen handling in routine laboratory diagnostics as well as in scientific studies to avoid preanalytical errors and, finally, misinterpretation of the data.

First molecular detection of Pneumocystis spp. in red foxes (Vulpes vulpeslinnaeus, 1758) and raccoon dogs (Nyctereutes procyonoidesgray, 1834).

Fungal organisms of the genus Pneumocystis may cause Pneumocystis pneumonia (PCP) in humans, but also domestic and wild mammals. Almost every animal species hosts its own genetically distinct Pneumocystis species, however information is sparse. In this study, 62 red foxes (Vulpes vulpes) and 37 raccoon dogs (Nyctereutes procyonoides) were collected in North-East Germany. The lung tissues of the animals were analysed by a new designed specific pan-Pneumocystis mtLSU rRNA gene PCR and sequencing. With this PCR, detection and discrimination of all known Pneumocystis spp. in a single step should be possible. This first detection of Pneumocystis spp. in 29/62 (46.8%) red foxes and 29/37 (78.4%) raccoon dogs indicated, that they harbour two dissimilar strains, as seen by specific single nucleotide position changes (SNPs). Nevertheless, five samples with contrary SNPs showed a probable inter-species transmission.

Effects of Different Oral Doses of Sodium Chloride on the Basal Acid-Base and Mineral Status of Exercising Horses Fed Low Amounts of Hay.

The provision of NaCl, according to current recommendations, to horses in moderate work has been shown to induce immediate postprandial acidosis. The present study aimed to clarify whether this NaCl induced acidosis i) persists beyond the immediate postprandial period, and ii) is still present after a 2 week adaptation period. Six adult warmblood mares in moderate work received daily 1.00 kg hay per 100 kg body weight (bwt) only together with 0.64 kg unprocessed cereal grains/100 kg bwt.d as fed basis. Using a 3x3 Latin Square, either 0 (NaCl-0), 50 (NaCl-50) or 100 (NaCl-100) g NaCl/d were fed together with the concentrates in two equal doses for 3 weeks. During the final week, a mineral digestibility trial was undertaken. The middle sodium and chloride intake (NaCl-50) at least met the most common recommendations for moderate work. Morning (7:00 AM) urine and venous blood samples were collected on days 0, 1-4, 8, and 15, and analysed for pH, acid-base status, creatinine and electrolyte concentrations. Fractional electrolyte clearances (FC) were determined. Mean apparent sodium digestibility ranged between 60-62% whereas chloride digestibility was consistently above 94%. Supplementing 100 g but not 50 g of NaCl resulted in significant reduction of blood pH and base excess as well as urinary pH and urine acid excretion. Both 50 g and 100 g NaCl supplementation caused a significant reduction in base and net acid-base excretion, urine density and potassium concentration, but increased urine sodium concentration and the FC of sodium and chloride (P < 0.05). This suggests that a high proportion of the recommended salt doses is excreted renally. The above effects of NaCl supplementation persisted over the 2 week measurement period. Results suggest that feeding 100 g NaCl to moderately exercising horses results in mild metabolic acidosis, whereas feeding 50 g according to current recommendations resulted in compensated acidosis.

Influenza A(H5N8) Virus Similar to Strain in Korea Causing Highly Pathogenic Avian Influenza in Germany.

Highly pathogenic avian influenza (H5N8) virus, like the recently described H5N8 strain from Korea, was detected in November 2014 in farmed turkeys and in a healthy common teal (Anas crecca) in northeastern Germany. Infected wild birds possibly introduced this virus.

Metabolic adaptations in the liver of born long-distance running mice.

PURPOSE: Long-distance runners have increased needs of energy supply. To unravel genetically based mechanisms required for efficient energy supply, we have analyzed hepatic metabolism of mice characterized by the inborn capacity to perform as long-distance runners. METHODS: The mouse model had been established by phenotypic selection for high treadmill performance for 90 generations and was characterized by approximately 3.8-fold higher running capacities (Dummerstorf high Treadmill Performance mouse line [DUhTP]) compared with unselected and also untrained controls (Dummerstorf Control mouse line [DUC]). From 7-wk-old male mice, serum and liver samples were collected and analyzed for messenger RNA, protein, and metabolite levels, respectively. RESULTS: In livers from DUhTP mice, we identified significantly higher messenger RNA transcript levels of peroxisome proliferator-activated receptor delta and higher protein levels of sirtuin-1, acetyl-CoA-synthetase, acetyl-CoA-carboxylase, phosphoenolpyruvate carboxykinase, and glutamate-dehydrogenase, suggesting higher gluconeogenesis and lipogenesis in DUhTP mice. In fact, higher hepatic levels of glycogen and triglycerides as well as higher concentrations of carbohydrate, fatty acid, and cholesterol metabolites were found in DUhTP mice. In parallel, in DUhTP mice, which did not have access to running wheels, a marked hyperlipidemia (cholesterol = 160% ± 8%, triglycerides = 174% ± 14% of controls, respectively), and abdominal obesity (DUhTP = 0.396 ± 0.019 g, DUC = 0.291 ± 0.019 g) were found. CONCLUSIONS: From our data, we conclude that the physiological basis of genetically fixed higher endurance-running performance in DUhTP marathon mouse is related to increased hepatic gluconeogenesis and lipogenesis. Expression of sirtuin 1 as well as of gluconeogenic and lipogenic key enzymes may be related to peroxisome proliferator-activated receptor delta. Metabolic adaptations presented in our study represent inborn features of superior endurance-running performance.