Occurrence of emerging ruminant viruses in goats in Poland.

Health status of Polish goat population in regard to the viral diseases remained mostly unknown. In order to determine serological status of Polish goats for selected emerging ruminant viruses, 365 serum samples collected between 2017 and 2019 in 36 districts within 10 of Polish provinces, were tested. No antibodies specific to Peste de Petite Ruminants Virus (PPRSV) and capripoxviruses (CaPV) were found in any of the tested animals. Only single individual (0.27%) was seropositive to Blutongue Virus (BTV). Antibodies directed to Schmallenberg Virus (SBV) were detected in 46 goats which represented 12.6% of the tested population. No association between seropositivity to SBV and year of sampling, province of origin, gender and age was found. In conclusion, among studied viral pathogens, currently only SBV seemed to be important for epidemiological status of Polish goats.

Antiobesity and Antioxidant Potentials of Selected Palestinian Medicinal Plants.

We evaluated the antioxidant and porcine pancreatic lipase inhibition (PPLI) activities of 90 plants extracts. The antioxidant activity was measured using the free-radical scavenging capacity (DPPH) and reducing power (RP) assays. The pancreatic lipase inhibition assay was used to determine the PPLI activity of plant extracts. Among the 90 plant extracts examined, 41.0 % crude extracts showed antilipase activity of more than 50%. The most active plants by means of IC50 value were Camellia sinensis (0.5 mg/ml), Ceratonia siliqua (leaves) (0.8 mg/mL), Curcuma longa (0.8 mg/mL), Sarcopoterium spinosum (1.2 mg/mL), and Mentha spicata (1.2 mg/mL). The antioxidant activity of plant extracts using the DPPH and RP assays reveals comparable results. The most active antioxidant extracts using both assays were the leaves and fruit epicarp of Rhus coriaria, areal parts of Sarcopoterium spinosum, and leaves of Ceratonia siliqua. Our results suggest natural resources that possess strong antioxidant and pancreatic lipase inhibitory activities with potential applications in the treatment and prevention of obesity and overweight. The extracts of Camellia sinensis, Ceratonia siliqua, Curcuma longa, Sarcopoterium spinosum, and Mentha spicata were proved to have a great potential as antioxidants and antiobesity agents.

Application of loop-mediated isothermal amplification (LAMP) assays for the detection of bovine herpesvirus 1.

Bovine herpesvirus-1 (BoHV-1), a causative agent of Infectious Bovine Rhinotracheitis (IBR), is responsible for high economic losses in cattle farming industry. The use of testing methods that allow early detection of BoHV-1-infected animals is a key element of each program of IBR eradication. The aim of the study was to design and evaluate two variants of LAMP isothermal tests with SYBR Green fluorescence probes, specific to the genes encoding gD and gE glycoproteins of BoHV-1. LAMP gE BoHV-1 assay was able to distinguish between gE- and gE+ strains of the virus. Both LAMP gD and gE assays were specific to BoHV-1 and did not react with other related to BoHV-1 alphaherpesviruses. Sensitivity of LAMP gD was 2x104 copies of the viral genome whereas for LAMP gE it was 2x105. Diagnostic sensitivity calculated for LAMP gD was 64.7% whereas for LAMP gE it was 80%. Diagnostic specificity for LAMP gD and LAMP gE was 78.9% and 89.3%, respectively. LAMP assay can be a rapid and simple method of diagnosis of acute BoHV-1 infections and discrimination of gE- strains. However, relatively low diagnostic sensitivity of the method can limit its use in routine diagnostics.

Replication kinetics of neuropathogenic and non-neuropathogenic equine herpesvirus type 1 (EHV-1) strains in primary murine neurons and ED cell line.

Equine herpesvirus type 1 (EHV-1) causes respiratory infections, abortion and neurological disorders in horses. Molecular epidemiology studies have demonstrated that a single-point mutation in DNA polymerase gene, resulting in an amino acid variation (N752/D752), is significantly associated with the neuropathogenic potential of EHV-1 strains. The aim of the study was to elucidate if there are any differences between neuropathogenic (EHV-1 26) and non-neuropathogenic (Jan-E and Rac-H) EHV-1 strains in their ability to infect neuronal cells. For the tested EHV-1 strains, cytopathic effect (CPE) was manifested by changed morphology of cells, destruction of actin cytoskeleton and nuclei degeneration, which led to focal degeneration. Moreover, EHV-1 26 strain caused fusion of the infected cells to form syncytia in culture. Real-time PCR analysis demonstrated that both neuropathogenic and non-neuropathogenic EHV-1 strains replicated in neurons and ED cells (equine dermal cell line) at a similar level. We can assume that a point mutation in the EHV-1 polymerase does not affect viral replication in this cell type.

Variability of non-structural proteins of equine arteritis virus during persistent infection of the stallion.

The genetic stability of ORF1a encoding non-structural proteins nsp1, nsp2, nsp3 and nsp4 of equine arteritis virus (EAV) has been analysed for nearly seven years in a persistently infected stallion of the Malopolska breed. Between November 2004 and June 2011, 11 semen samples were collected. Viral RNA extracted from semen of this carrier stallion was amplified, sequenced and compared with the sequences of the other known strains of EAV. Sequence analysis of ORF1a showed 84 synonymous and 16 non-synonymous mutations. The most variable part of ORF1a was the region encoding nsp2 protein with 13 non-synonymous substitutions. The degree of amino acid identity between isolates ranged from 98.91 to 100%. Only single non-synonymous mutations were detected in nsp1 (one substitution) and nsp4 (two substitutions). The most stable was nsp3 in which no amino acid substitutions were observed during the whole period of observation.

Prevalence of enterotoxin genes and antimicrobial resistance of coagulase-positive staphylococci recovered from raw cow milk.

Raw milk may be contaminated by enterotoxigenic coagulase-positive staphylococci (CPS). Several of these microorganisms show antimicrobial resistance, which poses a potential risk for consumers. The aim of this study was to determine the occurrence of enterotoxin genes and antimicrobial resistance of CPS isolated from cow milk. A total of 115 samples were analyzed for the presence of CPS according to the International Organization for Standardization standard (ISO 6888-2). The genes were identified using 2multiplex PCR assays. Resistance of the isolates to 10 antimicrobials was determined using the minimum inhibitory concentration method. Overall, 71 samples (62%) were contaminated with CPS and 69 isolates were further analyzed. Among them, 20 (29%) strains harbored the enterotoxin genes. The most commonly detected staphylococcal enterotoxin markers were sed, sej, and ser, whereas none of the analyzed isolates possessed the seb and see genes. Almost one-half of the tested strains (43%) were resistant to one or more antimicrobial agents. Resistance to penicillin was the most common, followed by sulfamethoxazole and chloramphenicol. On the other hand, all strains were susceptible to ciprofloxacin, erythromycin, gentamicin, cefoxitin, and streptomycin. None of the strains was positive for the mecA and mecC (methicillin-resistant Staphylococcus aureus) genes. These results indicate that enterotoxigenic and antimicrobial-resistant CPS strains are present in raw milk, which may be a potential risk for public health.

Rapid Detection of Bovine Respiratory Syncytial Virus in Poland Using a Human Patient-Side Diagnostic Assay.

Bovine respiratory syncytial virus (BRSV) plays a significant role in the etiopathogenesis of the respiratory syndrome in young cattle during their first year of life. Development of rapid and accurate BRSV diagnostic tools would aid in the appropriate control of this important pathogen. The objective of this study was to characterize infections induced by BRSV by means of rapid patient-side immunomigration assays used for diagnosis of human respiratory syncytial virus (hRSV) in humans. Nasal and tracheal swabs were obtained from healthy calves of various beef and dairy breeds - Holstein-Friesian, Simmental, Charolais, Belgian Blue and Limousin, between the ages of 5 and 12 months, from 26 farms. BRSV was identified using two rapid immunomigration assays, TruRSV® and Clearview® RSV, and compared with RT-PCR as a reference technique. BRSV was found in 73.1% of all the herds tested. High agreement with RT-PCR was obtained for TruRSV® (κ = 0.824), while in the case of the Clearview® RSV test, agreement with PCR was moderate (κ = 0.420). The results demonstrate that rapid patient-side immunomigration assays designed to detect hRSV can be used to accurately detect BRSV in field samples collected from cattle.

Hepatitis E virus antibody prevalence in wildlife in Poland.

Hepatitis E is an important public health problem mostly in developing but occasionally also in industrialized countries. Domestic and wildlife animals are considered reservoirs of the hepatitis E virus (HEV). Since no information on the prevalence of autochthonous HEV infections in human and animal in Poland is available, the aim of the study was to investigate the HEV seroprevalence of different wildlife species as potential virus reservoirs in the country. No HEV antibodies were found in any of the sera collected from the red deer (Cervus elaphus), European bison (Bison bonasus), roe deer (Capreolus capreolus), elk (Alces alces), fallow deer (Dama dama), sika deer (Cervus nippon), Tatra chamois (Rupicapra rupicapra tatrica) or brown bear (Ursus arctos). HEV-specific antibodies were detected in 44.4% (95% CI 38.3-50.7) serum samples originated only from wild boars. The percentage of seropositive wild boars differed significantly between the provinces and was positively correlated with the wild boar density and rurality of the area. This study showed that HEV circulates among wild boar population in Poland, and this species should be considered as an important reservoir of the virus.

Association between antibody status to bovine herpesvirus 1 and quality of milk in dairy herds in Poland.

Bovine herpesvirus 1 (BoHV1) is one of the most important pathogens of cattle; however, its effect on somatic cell count and milk components is not completely understood. The aim of the current study was to examine the effect of BoHV1 infection on quality of bovine bulk tank milk (BTM). A total of 1,790 individual blood samples collected at 28 dairy farms were used to determine the BoHV1 infection status of the herds with ELISA tests. The quality parameters of milk were evaluated by instrumental methods with BTM samples collected at monthly intervals from May 2011 to May 2012. The statistical analysis was performed to study the associations between BoHV1 herd status, quality of BTM, and herd-specific parameters. The risk factors influencing bulk milk somatic cell count (BMSCC) were estimated using the multivariable mixed-effects maximum likelihood regression model. The true prevalences of BoHV1 infection at the animal and herd levels were 49.3 and 64.6%, respectively. The average BMSCC differed significantly between the herds grouped accordingly to their BoHV1 infection status. Interestingly, the highest BMSCC was observed in the vaccinated herds (240.3×10(3) cells/mL). Additionally, the BoHV1 herd status had a significant effect on the fat content of BTM. The largest herds that were investigated had a BoHV1 seroprevalence over 30%. The herd status was considerably influenced by the numbers of cows in the herds. Besides, no significant differences in total bacterial count or protein content in milk from BoHV1-infected und uninfected herds were observed. An increase in BMSCC was observed during summer compared with the winter months regardless of the BoHV1 status of the herds. In the final multivariable regression model, the main risk factors associated with BMSCC were BoHV1 herd status, the percentage of BoHV1 infected animals in a herd, the number of cows in a herd, and the season. Our study suggests that BoHV1 infection may influence BMSCC levels, which are key parameters of BTM quality and a reference for subclinical mastitis in a herd. In conclusion, BoHV1 infection may cause economic losses by decrease both of quantity and quality of milk.

Application of real-time PCR for evaluation of distribution of equine herpesvirus type 1 in tissues of aborted fetuses.

A highly sensitive and specific real-time PCR assay was used for detection and quantitation of equine herpesvirus type 1 (EHV-1) in the different internal organs of aborted fetuses. Tissue samples from 23 aborted fetuses submitted to the Department of Virology of the National Veterinary Research Institute in Pulawy between 2012 and 2013 were used for testing. Total DNA was extracted using a phenol-chloroform-isoamyl alcohol standard protocol. A real-time PCR with forward and reverse primers encompassing a highly conserved region encoding viral glycoprotein B was adapted for diagnosis of EHV-1 infection. The detection limit of the assay was shown to be 6.0 × 10° of viral DNA copies and the obtained standard curve exhibited a linear range from 10° to 107 molecules. Sixteen out of twenty three aborted fetuses (69.5%) were positive for EHV-1 in real-time PCR. The highest EHV-1 DNA load was obtained for liver (mean Ct value: 15.7) and lung (18.2) samples, while the lowest was in the thymus (29.6) and placenta (28.4).

First report on equine herpesvirus type 4 isolation in Poland--evaluation of diagnostic tools.

Upper respiratory tract infections are still a serious problem in breeding and racing horses. The most common virological factors are EHV1 and EHV4, which are both a major cause of secondary infections. High EHV4 seroprevalence in Polish horses indicates a high transmission rate of this pathogen among horses and increases the need for proper diagnostics. The aim of this study was to develop a reliable laboratory diagnostic scheme for upper respiratory tract infections and to describe the first isolation of EHV4 in Poland. Twenty one nasal swabs collected from young horses under the age of 2 years showing clinical signs of equine rhinopneumonitis were tested with duplex PCR for simultaneous detection and differentiation between EHV1/EHV4. Positive samples were then subjected to virus isolation in Vero cells. Additionally, real-time PCR was developed which allowed viral copy numbers to be quantified and enabled defining that a DNA load below 10(3) copies per 1 ml of the sample reflected latent infection or decline of the disease. However, the sensitivity of traditional PCR proved to be sufficient in the diagnostic of the lytic infections and allowed identification of 10 EHV4 infected horses from which 3 strains were successfully isolated in cell culture. Another four EHV4 positive results were obtained by real-time PCR; but, a high Ct (threshold cycle) and a low virus DNA copy number suggested a latent infection. This report describes the first successful isolation of EHV4 from Polish horses.

Use of rapid human respiratory syncytial virus strip tests for detection of bovine respiratory syncytial virus in experimentally vaccinated calves.

Three different rapid strip tests: TRU RSV, BinaxNOW RSV and RSV Respi-strip were compared with RT-PCR and ELISA BRSV Ag for the ability to detect bovine respiratory syncytial virus (BRSV) in nasal swabs collected from calves experimentally vaccinated with live vaccine Rispoval RS-PI3. The reference strains of BRSV (375 and A51908) were detected by ELISA BRSV Ag whereas the strains of human respiratory syncytial virus (HRSV) and bovine parainfluenza virus type 3 (BPIV-3) were not. All rapid strip tests as well as RT-PCR reacted positively both to HRSV and BRSV reference strains and negatively to BPIV-3. The detection limit for RT-PCR was 39.1 TCID50 (strain 375 of BRSV), whereas for each of the rapid tests it was approximately 156 TCID50 and 312 TCID50 for antigen ELISA. Diagnostic sensitivity in detecting BRSV in nasal swabs for TRU RSV and RSV Respi-strip tests was 33% and 50% for BinaxNOW RSV. Diagnostic specificity of TRU RSV was 100%, whereas for both BinaxNOW and Respi-strip it was 87%. We concluded that TRU RSV could be used as a supportive rapid test for BRSV screening in nasal swabs taken directly on a farm. However, due to the small group of animals used in the experiment, the results should be regarded as preliminary and the study should be repeated on a larger number of animals.

Comparison of four RT-PCR assays for detection of bovine respiratory syncytial virus.

RT-PCR assays for detection of BRSV, based on four different sets of primers were optimized and evaluated for their sensitivity and specificity. Primers used in this study were specific for genes encoding three BRSV proteins, nucleoprotein N and glycoproteins F and G. Our results indicated that RT-PCR with primers B7:B8 for G protein was the most efficient in detecting BRSV. Starters B7:B8 reacted specifically only with BRSV strains, no cross-reaction with other closely related viruses to BRSV was observed. RT-PCR sensitivity was also high and amounted to 10(1.66) TCID50. Starters for F and N genes of BRSV were not sufficiently specific and cross-reacted with RNA of HRSV. RT-PCR with primers for the genes F and N of BRSV was characterized by a lower sensitivity than RT-PCR with primers B7:B8. In conclusion, RT-PCR specific to a sequence of glycoprotein G gene, seemed to be the most useful for BRSV detection.

Tuberculosis in a shopping centre, Portugal, 2004-5.

Genotyping enables to confirm or exclude a tuberculosis (TB) cluster. Excluding the link between cases is particularly important in countries with intermediate/high incidence of TB where the emergence of several TB cases in a particular location in space or time (higher than the expected) could be explained by chance alone. During 2004 and 2005, five TB cases occurred in five shops of a Portuguese shopping centre which employed a total of about 1000 workers. After an epidemiological survey, 52 close contacts were identified and screened. Latent tuberculosis infection was diagnosed in 10 contacts (eight family members and two work colleagues of cases). Genotyping of the Mycobacterium tuberculosis isolates revealed no link between the cases. For this reason no screening of all staff of the shopping centre was carried out. However, close contacts (52) and all fellow workers (1000) were kept under surveillance for two years, and no additional cases were diagnosed. The present analysis demonstrates that the exclusion of a chain of ongoing transmission by genotyping for the investigation of a cluster is cost-effective from the perspective of the public health service, because it allows to avoid unnecessary large scale screening operation and instead to direct resources to more effective measures of TB control.

Specific immunoprophylaxis of some viral diseases in cattle.

Specific immunoprophylaxis has played a key role in prevention and control of animal infectious diseases for many years. Vaccines are commonly used to diminish economic losses in animal production caused by infectious agents. Currently, respiratory diseases are the main health problem in cattle. BHV1, BRSV and BVD-MD viruses are among the most important pathogens worldwide. They can cause a disease themselves or they can be one of many agents causing a respiratory syndrome. The following article describes the specific immunoprophylaxis of cattle viral diseases caused by such viruses.

Insulin resistance of puberty in African-American children: lack of a compensatory increase in insulin secretion.

Type 2 diabetes has been increasing in children, mostly affecting minority populations at around the age of puberty. Despite a multitude of studies demonstrating pubertal insulin resistance/hyperinsulinemia in white children, data are almost non-existent in African-American children. The aim of the present study was to investigate the impact of puberty on glucose metabolism, insulin sensitivity and secretion in African-American children. Twenty prepubertal and 16 pubertal African-American subjects participated. All underwent a 3-h hyperinsulinemic (40 mU/m(2)/min) euglycemic clamp to determine insulin-stimulated glucose disposal, and a 2-h hyperglycemic (12.5 mmol/L) clamp to assess first- and second-phase insulin secretion. Body composition was assessed by dual energy X-ray absorptiometry (DEXA) and visceral and subcutaneous abdominal adiposity with computed tomography (CT) scan at L4-L5. Total glucose disposal, glucose oxidation and non-oxidative glucose disposal were significantly lower in the pubertal group compared with the prepubertal one (53.8 +/- 3.9 vs. 72.2 +/- 5.0 micromol/kg/min, p = 0.009; 23.3 +/- 1.1 vs. 31.6 +/- 1.7 micromol/kg/min, p = 0.001; and 30.0 +/- 3.3 vs. 40.5 +/- 3.9 micromol/kg/min, p = 0.049, respectively). Insulin sensitivity was approximately 30% lower in the adolescents compared with the prepubertal children. However, first- and second-phase insulin secretions were not different between the two groups (971.4 +/- 180.6 vs. 1044.0 +/- 191.4 pmol/L and 999.6 +/- 159.6 vs. 955.8 +/- 142.2 pmol/L, respectively). In conclusion, despite approximately 30% lower insulin sensitivity in African-American adolescents compared with prepubertal children, insulin secretion is not higher. This is in contrast to published findings in white children in whom insulin secretion is higher during puberty. These racial differences in physiologic adaptation to puberty could play a role in the higher prevalence of type 2 diabetes in African-American children at the time of puberty.

[Hospital discharge letter --Clinical information for the family physician]. / Nota de alta hospitalar--que informação clínica para o médico de família.

INTRODUCTION: There are aspects related with the proportion and quality of discharge letters which may influence the management of shared health care after hospital discharge; we describe them taking into consideration that no changes regarding the usual way of giving hospital discharge took place. AIMS: To determine the proportion of cases in which the hospital discharge of inpatients was accompanied with a discharge letter and to measure its quality. METHODS: A descriptive and transverse observation study referring to the year of 1996. The authors observed, in the clinical files of this Department, the total cases of discharge of inpatients with ischaemic heart disease (123 cases). Besides measuring the variable "discharge letter existence", we studied the composed variable discharge letter quality (existence of clinical history data, objective observation data, complementary test results, diagnosis, therapeutic procedures, and also the readability of the clinical information and the existence of the legible name of the specialist). RESULTS: From the study of 121 cases of discharge of inpatients with ischaemic heart disease, 90.2% presented discharge letters. When studied, their quality was considered sufficient in 57.7% and insufficient in the rest of the cases. We point out that 29.7% of the discharge letters did not present objective observation data. DISCUSSION: The clinical information provided in this Department, at the time of hospital discharge, presents a higher proportion and quality than that observed in other studies.

[Murine typhus in Portugal]. / Tifo murino em Portugal.

Murine typhus or endemic typhus is a wide spread infectious disease, with a low prevalence in developed countries, but surely underdiagnosed. Its relative benignity, the similarity to other infectious diseases and the discretion of its epidemiologic chain, as well as the usual unexpectedness of its existence, makes its diagnosis more difficult. The clinical presentation and evolution of this case illustrate the necessity of being aware of this nosological entity.

An international validation of the interaction with disabled persons scale.

The Interaction with Disabled Persons Scale (IDP) was devised to measure attitudes in terms of discomfort reported about social interaction with people with disabilities. The Scale has been used in Australia for ten years. This article reports results of an international validation project that was designed to determine whether psychometric characteristics and norms emerging for Australian groups apply elsewhere. A methodological proforma was developed to maximize uniformity of data collection across nine countries: Australia, Canada, Croatia, England, Germany, Hong Kong, Poland, Scotland and the United States. In most countries the Scale was administered in English; however it also was translated into Germany, Polish, French and Croat. Results indicate that across countries mean scores fell within ten points, similar moderate to high levels of item homogeneity occurred and level of prior contact with people with disabilities emerged as the strongest predictor of IDP scores. It was concluded that the IDP Scale is a valid measure that is able to discriminate between respondents within the countries included in the study.

Specificity and nucleotyping studies of a gp50-based polymerase chain reaction assay for detection of pseudorabies virus.

PCR primers that amplify a region of the gp50 envelope glycoprotein gene of a number of vaccinal and field strains of pseudorabies virus (PRV) have been previously described (Galeota-Wheeler and Osorio, Am J Vet Res 1991: 52; 1799-1803). This gp50-based PCR assay was tested for its diagnostic applicability, utilizing a panel of nine PRV isolates and 13 related herpesviruses, originating from domestic animal species and man. Slight modifications to the original PRV PCR protocol ensured that false positive PCR products from avian herpesviruses were not evident in agarose gel electrophoresis analysis. Nucleotide sequence data derived from the PCR product revealed that the region of the genome amplified was markedly conserved and allowed only for virus subgrouping, rather than definitive isolate characterization.