Road crossings change functional diversity and trait composition of stream-dwelling macroinvertebrate assemblages.

Functional diversity is regarded as a key concept in understanding the link between ecosystem function and biodiversity, and is therefore widely investigated in relation to human-induced impacts. However, information on how the intersection of roads and streams (hereafter road crossings, representing a widespread habitat transformation in relation to human development), influences the functional diversity of stream-dwelling macroinvertebrates is still missing. The general aim of our study was to provide a comprehensible picture on the impacts of road crossing structures on multiple facets of the functional diversity of stream-dwelling macroinvertebrates. In addition, we also investigated changes in trait structure. Our research showed that road crossing structures had negative impacts on functional richness and dispersion; i.e., functional diversification. However, we found no significant impact on functional divergence and evenness components. We found a decrease in functional redundancy at road crossing structures. This indicates a reduced ability of the community to recover from disturbances. Finally, we found that road crossings drive stream habitat and hydrological changes in parallel with modification of the trait composition of stream-dwelling macroinvertebrate assemblages. All these results suggest that road crossings cause notable changes in the functional diversity of stream-dwelling macroinvertebrate assemblages.

Elimination of porcine reproductive and respiratory syndrome virus infection using an inactivated vaccine in combination with a roll-over method in a Hungarian large-scale pig herd.

BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) causes severe economic losses worldwide and only four countries in Europe are free from PRRSV. Complete depopulation-repopulation is the safest and fastest, but also the most expensive method for eradicating PRRSV from a population. Another possible way to eliminate an endemic PRRSV infection is to replace the infected breeding stock by gilts reared isolated and protected from PRRSV on an infected farm. With this method it is possible to maintain continuous production on the farm. The authors report the first successful elimination of PRRSV in a Hungarian large-scale pig farm by using an inactivated vaccine and performing segregated rearing of the offspring. CASE PRESENTATION: The study was performed on a PRRSV infected farm (Farm A) with 1475 sows. The clinical signs of reproductive failure had been eliminated previously by using an inactivated vaccine (Progressis®, Ceva). At the beginning of the elimination programme, gilts intended for breeding were vaccinated at 60 and 90-100 days of age. After that, gilts selected for breeding were vaccinated at 6 months of age, on the 60-70th day of pregnancy and at weaning. Approximately 1200 piglets from vaccinated sows were transported at 7 weeks of age to a closed, empty farm (Farm B) after being tested negative for PRRSV by a polymerase chain reaction (PCR) method, and then were reared here until 14 weeks of age. At this age, all pigs were tested by PRRS ELISA. Seronegative gilts (n = 901) were subsequently transported from Farm B to a third, closed and empty farm (Farm C), and (having reached the breeding age) they were inseminated here after a second negative serological test (ELISA). At the same time, Farm A was depopulated, cleaned and disinfected. All pregnant gilts were transported from Farm C to Farm A after being re-tested negative for antibodies against PRRSV. Follow-up serology tests were performed after farrowing and results yielded only seronegative animals. Based on the subsequent negative test results, the herd was declared PRRSV free by the competent authority. CONCLUSIONS: The presented farm was the first during the National PRRS Eradication Programme of Hungary to eradicate PRRSV successfully by vaccinating the sows with an inactivated vaccine and performing segregated rearing of the offspring. Production was almost continuous during the whole process of population replacement.

Fluorescence-Based Real-Time Analysis of Osteoclast Development.

Osteoclasts are multinucleated cells of hematopoietic origin which are critically involved in physiological and pathological bone resorption. They develop from myeloid progenitors through characteristic gene expression changes and intercellular fusion. This process is directed by M-CSF and RANKL which are also able to trigger osteoclast development from bone marrow cells in vitro. Osteoclasts are conventionally visualized by histochemical staining followed by manual counting, which hinders kinetic studies and automated quantification. Here we describe two fluorescence-based assays for the real-time analysis of myeloid cell to osteoclast development (FRAMCO) in primary mouse bone marrow cell cultures. Both assays rely on red-to-green fluorescence conversion of the membrane-targeted tdTomato/membrane-targeted eGFP (mTmG) transgene by Cre recombinase driven by the osteoclast-specific cathepsin K promoter (Ctsk-Cre). In the first assay (FRAMCO1.1), osteoclast-specific gene expression triggers red-to-green color conversion of cells carrying both the Ctsk-Cre and mTmG transgenes. In the second assay (FRAMCO1.2), red-to-green fluorescence conversion is triggered by fusion of neighboring co-cultured bone marrow cells separately carrying either the Ctsk-Cre or the mTmG transgenes. The two assays were tested using a high-content confocal fluorescence imaging system, followed by automated quantification. The FRAMCO1.1 assay showed robust red-to-green fluorescence conversion of more than 50% of the culture (including mononuclear cells) within 3 days under osteoclastogenic conditions. The FRAMCO1.2 assay showed a less robust but still readily measurable red-to-green color conversion in multinuclear cells within 5 days of differentiation. The assays required both the Ctsk-Cre and the mTmG transgenes and gave no signals in parallel macrophage cultures. The proper functioning of the two assays was also confirmed at the DNA, mRNA and bulk protein level. The assay systems were validated using lisophosphatidylcholine, a previously reported inhibitor of preosteoclast fusion. Taken together, our assays allow high-throughput automated real-time analysis of two critical aspects of osteoclast development, facilitating the screening for novel drug candidates for the pharmacological control of osteoclast-mediated bone resorption.

Chronic Oral Selegiline Treatment Mitigates Age-Related Hearing Loss in BALB/c Mice.

Age-related hearing loss (ARHL), a sensorineural hearing loss of multifactorial origin, increases its prevalence in aging societies. Besides hearing aids and cochlear implants, there is no FDA approved efficient pharmacotherapy to either cure or prevent ARHL. We hypothesized that selegiline, an antiparkinsonian drug, could be a promising candidate for the treatment due to its complex neuroprotective, antioxidant, antiapoptotic, and dopaminergic neurotransmission enhancing effects. We monitored by repeated Auditory Brainstem Response (ABR) measurements the effect of chronic per os selegiline administration on the hearing function in BALB/c and DBA/2J mice, which strains exhibit moderate and rapid progressive high frequency hearing loss, respectively. The treatments were started at 1 month of age and lasted until almost a year and 5 months of age, respectively. In BALB/c mice, 4 mg/kg selegiline significantly mitigated the progression of ARHL at higher frequencies. Used in a wide dose range (0.15-45 mg/kg), selegiline had no effect in DBA/2J mice. Our results suggest that selegiline can partially preserve the hearing in certain forms of ARHL by alleviating its development. It might also be otoprotective in other mammals or humans.

Effects of Long-Term Selection in the Border Collie Dog Breed: Inbreeding Purge of Canine Hip and Elbow Dysplasia.

Pedigree data of 13,339 border collie dog was collected along with canine hip dysplasia (CHD) and canine elbow dysplasia (CED) records (1352 CHD and 524 CED), and an inbreeding-purging (IP) model was created. Ancestral inbreeding coefficients were calculated by using a gene dropping simulation method with GRain 2.2 software. Cumulative logit models (CLM) for CHD and CED were fitted using a logit-link Poisson distribution and the classical (F_W), and ancestral inbreeding (F_BAL, F_KAL, and F_KAL_NEW) coefficients as linear regression coefficients. The effective population size was calculated from F_W and decreased in the examined period along with an increase of F_W; however, slight differences were found as a consequence of breeding dog imports. CHD values were lowered by the expansion of F_BAL, as the alleles had been inbred in the past. For CHD, signs of purging were obtained. There was a positive trend regarding the breeding activity (both sire and dam of the future litters should be screened and certified free from CHD and CED), as years of selection increased the frequency of alleles with favorable hip and elbow conformation. Division of the ancestral inbreeding coefficient showed that alleles that had been identical by descent (IBD) for the first time (F_KAL_NEW) had a negative effect on both traits, while F_KAL has shown favorable results for alleles IBD in past generations. Some authors had proven this phenomenon in captive populations or experimental conditions; however, no evidence of inbreeding purge has ever been described in dog populations. Despite the various breeding practices, it seems that alleles of these polygenic disorders could be successfully purged out of the population with long-term selection.

Anti-PD-1 Therapy Does Not Influence Hearing Ability in the Most Sensitive Frequency Range, but Mitigates Outer Hair Cell Loss in the Basal Cochlear Region.

The administration of immune checkpoint inhibitors (ICIs) often leads to immune-related adverse events. However, their effect on auditory function is largely unexplored. Thorough preclinical studies have not been published yet, only sporadic cases and pharmacovigilance reports suggest their significance. Here we investigated the effect of anti-PD-1 antibody treatment (4 weeks, intraperitoneally, 200 µg/mouse, 3 times/week) on hearing function and cochlear morphology in C57BL/6J mice. ICI treatment did not influence the hearing thresholds in click or tone burst stimuli at 4-32 kHz frequencies measured by auditory brainstem response. The number and morphology of spiral ganglion neurons were unaltered in all cochlear turns. The apical-middle turns (<32 kHz) showed preservation of the inner and outer hair cells (OHCs), whilst ICI treatment mitigated the age-related loss of OHCs in the basal turn (>32 kHz). The number of Iba1-positive macrophages has also increased moderately in this high frequency region. We conclude that a 4-week long ICI treatment does not affect functional and morphological integrity of the inner ear in the most relevant hearing range (4-32 kHz; apical-middle turns), but a noticeable preservation of OHCs and an increase in macrophage activity appeared in the >32 kHz basal part of the cochlea.

Inbreeding depression for kit survival at birth in a rabbit population under long-term selection.

BACKGROUND: Accumulation of detrimental mutations in small populations leads to inbreeding depression of fitness traits and a higher frequency of genetic defects, thus increasing risk of extinction. Our objective was to quantify the magnitude of inbreeding depression for survival at birth, in a closed rabbit population under long-term selection. METHODS: We used an information theory-based approach and multi-model inference to estimate inbreeding depression and its purging with respect to the trait 'kit survival at birth' over a 25-year period in a closed population of Pannon White rabbits, by analysing 22,718 kindling records. Generalised linear mixed models based on the logit link function were applied, which take polygenic random effects into account. RESULTS: Our results indicated that inbreeding depression occurred during the period 1992-1997, based on significant estimates of the z-standardised classical inbreeding coefficient z.FL (CI95% - 0.12 to - 0.03) and of the new inbreeding coefficient of the litter z.FNEWL (CI95% - 0.13 to - 0.04) as well as a 59.2% reduction in contributing founders. Inbreeding depression disappeared during the periods 1997-2007 and 2007-2017. For the period 1992-1997, the best model resulted in a significantly negative standardised estimate of the new inbreeding coefficient of the litter and a significantly positive standardised estimate of Kalinowski's ancestral inbreeding coefficient of the litter (CI95% 0.01 to 0.17), which indicated purging of detrimental load. Kindling season and parity had effects on survival at birth that differed across the three periods of 1992-1997, 1997-2007 and 2007-2017. CONCLUSIONS: Our results support the existence of inbreeding depression and its purging with respect to kit survival at birth in this Pannon White rabbit population. However, we were unable to exclude possible confounding from the effects of parity and potentially other environmental factors during the study period, thus our results need to be extended and confirmed in other populations.

[The impact of a checklist on the short-term complications of airway management in adults]. / Ellenorzo lista hatása a légútbiztosítás korai szövodményeire felnottekben.

Introduction: Airway management is an integral part of general anaesthesia, which may lead to severe short- and long-term complications. Aim: We assessed whether the application of a checklist for the steps of airway management reduces the number of complications in our institute. Method: In our observational, prospective, controlled study we made a checklist and a data collection sheet. Each airway management was performed for one month without the checklist and then for one month in the possession of the checklist. We evaluated the outcome of airway maneuvers and the occurrence of related early complications before and after the introduction of the checklist. The primary endpoint was the incidence of unexpected difficult airway. The secondary endpoints were difficult intubation, successful first intubation, aspiration, cardiac arrest, post-induction hypotension and desaturation, soft tissues/teeth injuries. Our results were also corrected for factors that affect the risk of complications (urgency of interventions, medical experience). Results: We did not find any difference in the frequency of acute complications before the introduction of the checklist (n = 439) and during the subsequent period (n = 423). At the primary endpoint (7.29% and 6.14%), there was no substantive difference (1.15%, 95% CI: -2.26%-4.56%, p = 0.5). No differences were found regarding the secondary and other endpoints. Following the correction of risk factors, there was no impact of the checklist on the incidence of complications. Conclusion: The introduction of the checklist in itself did not result in a significant change in the risk of short-term complications of airway management in our institution. Orv Hetil. 2019; 160(26): 1025-1035.

Comparison of various advanced oxidation processes for the degradation of phenylurea herbicides.

Various types of advanced oxidation processes (AOPs), such as UV photolysis, ozonation, heterogeneous photocatalysis and their combinations were comparatively examined at the same energy input in a home-made reactor. The oxidative transformations of the phenylurea herbicides fenuron, monuron and diuron were investigated. The initial rates of transformation demonstrated that UV photolysis was highly efficient in the cases of diuron and monuron. Ozonation proved to be much more effective in the transformation of fenuron than in those of the chlorine containing monuron and diuron. In heterogeneous photocatalysis, the rate of decomposition decreased with increase of the number of chlorine atoms in the target molecule. Addition of ozone to UV-irradiated solutions and/or TiO2-containing suspensions markedly increased the initial rates of degradation. Dehalogenation of monuron and diuron showed that each of these procedures is suitable for the simultaneous removal of chlorinated pesticides and their chlorinated intermediates. Heterogeneous photocatalysis was found to be effective in the mineralization.

A 4-year study of a natural tick-borne encephalitis virus focus in Hungary, 2010-2013.

A tick-borne encephalitis virus focus was identified in a former goat pasture that had been associated with a milk-borne encephalitis outbreak in 2007. Ticks and rodents were sampled monthly from April 2010 to October 2013 on two separate 0.5 ha sampling sites. At site 1, three tick-borne encephalitis virus strains were isolated from a total of 7,247 sampled ticks; 28 of the 539 tested sera (5.19%) were seropositive. At site 2, from the 2,369 sampled ticks, virus was not isolated, tests of 284 rodent sera resulted in 14 positives (4.93%). For survival, the virus needs a territory with continuously dense rodent and tick population, although observed TBEV prevalence was low both in ticks and in rodents. Sampling points of positive ticks and rodents did not coincided exactly, at a certain time only some m(2) territory is dangerous, these hot spots change unpredictably as positive ticks die or move on with their hosts.

European rodent on the edge: status and distribution of the Vojvodina blind mole rat.

Recent research of blind mole rats of the species complex Nannospalax (superspecies leucodon) identified a small and fragmented population of these rodents on both sides of the Hungarian-Serbian border. Cytogenetic investigations proved that this population karyologically identical with the Vojvodina blind mole rat described earlier as Nannospalax (leucodon) montanosyrmiensis. Based on cytochrome b gene sequences obtained from three specimens originating from separate locations, these blind mole rats form a discrete phylogenetic clade which, with a difference of about 10%, is well separated from other blind mole rat taxa inhabiting the Carpathian Basin. The taxon has only two extant populations that are 150 km apart from each other. The combined occupied area is estimated to be less than 10 km(2), and the total estimated number of individuals is less than 300. These two remaining populations are heavily fragmented and under imminent threat by the establishment of tree plantations, small-scale and agro-industrial farms and land development. The situation is further aggravated by the fact that 80% of the individuals inhabit unprotected areas. A study of the landscape history of the wider area surrounding one of the populations - based on military maps spanning over the last 200 years - has shown a drastic decrease in the extent and quality of potential habitats. Based on our present knowledge, the Vojvodina blind mole rat is one of the most seriously threatened, rarest mammal in Europe, the remaining population of which can be wiped out within years unless immediate conservation action is taken.