Western blot (Wb) is considered to be the gold standard test for Trichinella infection serology, since this method allows specific Trichinella antigens to be distinguished from cross-reactive ...antigens. This is not the case with widely used antibody assay techniques - indirect immunofluorescence and ELISA - which are sensitive, but subject to crossreactions that make the interpretation of weakly positive results difficult. Application of Trichinella spiralis muscle larvae excretory-secretory (ES) antigens for the specific antibody detection in ELISA resulted in improved specificity compared to that of crude worm extract that was previously in use, but since production of ES has not yet been standardized, differences among laboratories occur. For this reason, the Wb profile of serum samples from different T. spiralis infected host species: human, horse, swine and dog, was investigated in the Serbian National Reference Laboratory for Trichinellosis (NRLT). The common feature of the obtained Wb profiles was the appearance of a triad of bands with molecular masses (Mw) of 45, 49, and 53 kDa. The very same triad was recognized by a monoclonal antibody (mAb) 7C2C5 specific for an immunodominant epitope unique to the muscle larvae stage of all species in the genus Trichinella. Inhibition studies confirmed that mAb and anti-Trichinella antibodies from sera competed for the same parasite epitope. Based on the obtained results, the NRLT introduced the recognition of the above mention triad as the basis for specific anti-Trichinella antibodies detection in the sera of infected hosts.
In this work we investigated presence of Trichinella species at endemic districts Branicevo and Podunavlje in population of domestic and wild animals. We examined 211 samples of muscular tissue from ...domestic pigs and 30 samples from dogs, by artificial digestion. Positive result was in 25 (11, 85%) samples from pigs and 7 (23,33%) dogs. Significant different (p<0, 01) was in percentage of infection in population of pigs from backyards and pigs from industrial farms. Among wild life, 338 samples were examined. Trichinella infections were detected in 7 from 174 (4,02%) wild boars (Sus scrofa), in 8 from 118 (6, 78%) foxes (Vulpes vulpes), in 8 from 42 (19, 05%) jackals (Canis aureus) and in all for examined wolves (Canis lupus). During serology survey, performed at industrial farms in 150 samples of swine sera we didn’t detect anti-Trichinella antibodies by ELISA test. In backyards we confirmed positive results in 26 (12, 68%) swine sera, weakly positive results in 6 (2, 93%) sera samples and 13 (6, 34%) negative results. From 11 serum samples of stray dogs from municipality of Pozarevac, 8 have specific anti-Trichinella antibodies, 1 was suspect and 2 were negative. We examined 17 samples of meet juice from wild boars when 7 samples were positive and in 10 samples we didn’t detect anti-Trichinella antibodies by ELISA test. Results from serology investigation were confirmed by artificial digestion. All positive and weakly positive samples in ELISA test (32 samples) were confirmed as positive in Western blot test. Samples examined by Multiplex PCR were from 6 animal species: 25 domestic swine, 7 wild boars, 8 foxes, 8 jackals, 4 wolves and 7 dogs. We obtained result from 53 samples and non result in 6 (10, 2%) samples. Trichinella spiralis were identified in 43 (72, 9%) samples and Trichinella britovi in 7 samples (11,8%). Mix infection with boat species were in 3 samples (5, 1%). Trichinella spiralis was found in 25 domestic pigs, 5 dogs, 5 foxes, 5 jackals and 3 wild boars. Trichinella britovi was found only in wild animals (1 wild boar, 1 jackal, 1 fox and 4 wolves). Mix infection (T. spiralis ( T. britovi) was found in 2 foxes and 1 jackal. In population of domestic pig and dogs only T. spiralis was found. Geographical distribution showed at Podunavlje district only presents of T. spiralis and at Branicevo district presents of T. spiralis i T. britovi. With epizootiologial and geographical data, identification of Trichinella species in population of domestic and wild animals at Branicevo and Podunavlje district we obtained data about location (latitude/longitude), identified points of interest (industrial farms, hunting section) as potential source for infection. During this investigation, for the first time mix infection with T. spiralis and T. britovi was detected in wild animals, which point out possible circulation between wild and domestic cycles and potential risk for domestic animals. With results of Western blot and Multiplex PCR investigation this work has significant contribution for epizootiological surveillance of Trichinella infection in domestic and wild animals.
U ovom radu ispitivana je prisutnost trihinela na endemskom području Braničeva i Podunavlja kod domaćih i divljih životinja, kao i identifikacija prisutnih vrsta roda Trichinella. Metodom veštačke digestije ispitano je 211 uzoraka poreklom od domaćih svinja i pozitivan nalaz je dobijen kod 25 (11,85%) uzoraka. Ustanovljena je signifikantna razlika (p<0,01) u procentu pojavljivanja trihineloze kod domaćih svinja poreklom sa individualnih gazdinstava i industrijskih farmi. Ispitivanjem je obuhvaćeno i 338 uzoraka poreklom od divljih životinja i 30 pasa. Infekcija je utvrđena kod 7 od 174 (4,02%) ispitanih divljih svinja (Sus scrofa), kod 8 od 118 (6,78%) lisica (Vulpes vulpes), kod 8 od 42 (19,05%) šakala (Canis aureus), kod sva 4 (100%) pregledana vuka (Canis lupus) i 7 (23,33%) pasa (Canis familiaris). Imunoenzimskim testom (ELISA test) na 150 uzoraka krvnih seruma domaćih svinja poreklom sa farmi nije utvrđeno prisustvo specifičnih antitela protiv vrsta roda Trichinella. Na individualnim gazdinstvima ustanovljeno je 26 pozitivnih (12,68%) uzoraka krvnih seruma svinja, 6 (2,93%) je bilo slabo pozitivno, a 13 (6,34%) negativno. Od ukupno 11 pregledanih uzoraka krvi pasa lutalica koji su poticali sa teritorije opštine Požarevac 8 uzoraka je reagovalo pozitivno, 1 slabo pozitivno i 2 su reagovala negativno. Imunoenzimski test (ELISA test) na uzorcima mesnog soka divljih životinja urađen je na 17 uzoraka, od čega je 7 bilo serološki pozitivno, dok kod 10 uzoraka nije ustanovljeno prisustvo antitela protiv Trichinella spp. Rezultati seroloških ispitivanja su bili podudarni sa rezultatima veštačke digestije. Imunoblot metodom je ispitano 36 uzoraka krvnih seruma domaćih svinja. Svi uzorci koji su pokazali pozitivan i slabo pozitivan rezultat u ELISA testu (32 uzorka) bili su pozitivni na Western blot testu. Uzorci ispitani multiplex PCR tehnikom su poticali od 6 vrsta životinja i to: 25 domaćih svinja, 7 divljih svinja, 8 lisica, 8 šakala, 4 vuka i 7 pasa. Rezultati su dobijeni za 53 uzoraka, dok kod ukupno 6 uzoraka identifikacija nije uspela (10,2%) i test PCR je proglašavan negativan. Trichinella spiralis je identifikovana u 43 uzoraka (72,9%). Trichinella britovi je identifikovana u 7 uzoraka (11,8%). Prisustvo obe vrste T. spiralis i T. britovi utvrđeno kod 3 uzorka (5,1%). Trichinella spiralis je utvrđena kod 25 domaćih svinja, 5 pasa, 5 lisica, 5 šakala i 3 divlje svinje. Trichinella britovi je utvrđena samo kod divljih životinja i to 1 divlje svinje, 1 šakala, 1 lisice i 4 vuka. Prisustvo mešovite infekcije (T. spiralis i T. britovi) utvrđeno je kod 2 lisice i 1 šakala. U populaciji domaćih svinja i pasa lutalica utvrđeno je prisustvo samo T. spiralis. U odnosu na geografsku rasprostranjenost na teritoriji Podunavskog okruga utvrđeno je prisustvo T. spiralis, dok je na teritoriji Braničevskog okruga utvrđeno prisustvo T. spiralis i T. britovi. Na osnovu sprovedenih istraživanja i dobijenih rezultata za geografsko pozicioniranje utvrđenih slučajeva trihineloze i identifikovanih vrsta roda Trichinella kod domaćih i divljih životinja na teritoriji epizootiološkog područja Braničevskog i Podunavskog okruga, dobijeni su podaci o geografskim koordinatama (dužina/širina), lokacije, a posebno tačke od interesa (industrijske farme, lovišta), kao potencijalnih izvora infekcije za prijemčive vrste životinja. Tokom ovog istraživanja po prvi put je dokazana mešovita infekcija sa T. spiralis i T. britovi kod divljih životinja, što ukazuje na mešanje domaćeg i silvatičnog ciklusa i ukazuje na potencijalni rizik za domaće vrste životinja. Uz to, rezultati dobijeni primenom imunoblota i multiplex PCR metode predstavljaju značajan doprinos epizootiološkom nadzoru infekcije vrstama roda Trichinella kod domaćih i divljih životinja.
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► A large number of sera samples from humans and pigs were tested. ► A distinctive Western blot pattern of Trichinella infections is defined. ► The Western blot pattern unequivocally ...distinguishes true positive from false positive sera.
Trichinellosis is a zoonotic disease caused by parasites of the genus Trichinella, which have a cosmopolitan distribution. For diagnostic purposes, a confirmatory test for ELISA-positive human and pig sera such as Western blotting is required, due to the high number of ELISA false positive sera. The objective of this study was to identify the Trichinella-specific antigens most frequently recognized by sera from Trichinella-infected humans and pigs, so as to define a distinctive pattern of Trichinella infection in sera from infected hosts using Western blots which allow false positive sera to be distinguished from true positive sera. Using excretory/secretory antigens, 450 human sera were tested by Western blotting: 150 from persons with a confirmed diagnosis of trichinellosis and 300 from persons who did not have trichinellosis but who tested positive by ELISA (i.e., false positives). We also tested 210 pig sera: (i) 30 from pigs experimentally infected with Trichinella spiralis; (ii) 90 from naturally T. spiralis-infected pigs; and (iii) 90 from pigs not infected with Trichinella, as shown after artificial digestion of the diaphragm pillars, yet which tested positive by ELISA (i.e., false positives). All true positive sera (i.e., sera from persons with confirmed trichinellosis as well as sera from naturally and experimentally infected pigs), reacted with a three-band pattern ranging in size from 48–72kDa. A distinctive pattern for recognizing Trichinella spp. infections in humans and pigs by Western blots is defined; it shows a sensitivity of 100% and it allows sera from Trichinella-infected humans and pigs to be distinguished from sera from persons and pigs that were not infected with Trichinella spp. (100% specificity).
Effects of the application of the trichinellosis control program in the endemic part of Serbia Tešić, M. (Faculty of Veterinary Medicine, Belgrade (Serbia)); Teodorović, V. (Faculty of Veterinary Medicine, Belgrade (Serbia)); Mirilović, M. (Faculty of Veterinary Medicine, Belgrade (Serbia)) ...
Proceedings of the International Conference Biological Food Safety and Quality,
2012
Conference Proceeding
Trichinellosis is a disease of humans and animals caused by the parasite that belongs to the family Trichinellidae, genus Trichinella, and infection with this parasite has been described in over 100 ...species of mammals in all continents, while the most known Trichinella species in Serbia is Trichinella spiralis. It has high infectivity and pathogenicity in humans, and infection of humans is caused by consumption of infected meat. In the investigated region trichinellosis has been present for many years. Local authorities have conducted partial pest control, but even when positive, results have shown the percentage of infected pigs is still high, and especially in individual slaughter pigs (0.6024 and 0.3865 % positive in 2004 and 2005, respectively). In that sense, the relevant state authorties decided to implement systematic control and suppression of trichinellosis in that region. Besides the zoonotic characteristics of trichinellosis, without any doubt it is also an economic problem, which indicates the necessity of drafting a disease control program for the observed region. During preparation of a consistent disease control and eradication program, the starting point was legislation in place and determination of the necessary inputs and outputs for it. Justification for drafting the program was demonstrated after application of the cost-benefit analysis (CBA), while the evaluation of the program was conducted based on the net present value (NPV) and ratio of the benefit and cost (B/C).
Effects of the application of the trichinellosis control program in the endemic part of Serbia Tešić, M. (Faculty of Veterinary Medicine, Belgrade (Serbia)); Teodorović, V. (Faculty of Veterinary Medicine, Belgrade (Serbia)); Mirilović, M. (Faculty of Veterinary Medicine, Belgrade (Serbia)) ...
Proceedings of the International Conference Biological Food Safety and Quality,
2012
Conference Proceeding
Trichinellosis is a disease of humans and animals caused by the parasite that belongs to the family Trichinellidae, genus Trichinella, and infection with this parasite has been described in over 100 ...species of mammals in all continents, while the most known Trichinella species in Serbia is Trichinella spiralis. It has high infectivity and pathogenicity in humans, and infection of humans is caused by consumption of infected meat. In the investigated region trichinellosis has been present for many years. Local authorities have conducted partial pest control, but even when positive, results have shown the percentage of infected pigs is still high, and especially in individual slaughter pigs (0.6024 and 0.3865 % positive in 2004 and 2005, respectively). In that sense, the relevant state authorties decided to implement systematic control and suppression of trichinellosis in that region. Besides the zoonotic characteristics of trichinellosis, without any doubt it is also an economic problem, which indicates the necessity of drafting a disease control program for the observed region. During preparation of a consistent disease control and eradication program, the starting point was legislation in place and determination of the necessary inputs and outputs for it. Justification for drafting the program was demonstrated after application of the cost-benefit analysis (CBA), while the evaluation of the program was conducted based on the net present value (NPV) and ratio of the benefit and cost (B/C).
Effects of the application of the trichinellosis control program in the endemic part of Serbia Tešić, M. (Faculty of Veterinary Medicine, Belgrade (Serbia)); Teodorović, V. (Faculty of Veterinary Medicine, Belgrade (Serbia)); Mirilović, M. (Faculty of Veterinary Medicine, Belgrade (Serbia)) ...
Proceedings of the International Conference Biological Food Safety and Quality,
2012
Conference Proceeding
Trichinellosis is a disease of humans and animals caused by the parasite that belongs to the family Trichinellidae, genus Trichinella, and infection with this parasite has been described in over 100 ...species of mammals in all continents, while the most known Trichinella species in Serbia is Trichinella spiralis. It has high infectivity and pathogenicity in humans, and infection of humans is caused by consumption of infected meat. In the investigated region trichinellosis has been present for many years. Local authorities have conducted partial pest control, but even when positive, results have shown the percentage of infected pigs is still high, and especially in individual slaughter pigs (0.6024 and 0.3865 % positive in 2004 and 2005, respectively). In that sense, the relevant state authorties decided to implement systematic control and suppression of trichinellosis in that region. Besides the zoonotic characteristics of trichinellosis, without any doubt it is also an economic problem, which indicates the necessity of drafting a disease control program for the observed region. During preparation of a consistent disease control and eradication program, the starting point was legislation in place and determination of the necessary inputs and outputs for it. Justification for drafting the program was demonstrated after application of the cost-benefit analysis (CBA), while the evaluation of the program was conducted based on the net present value (NPV) and ratio of the benefit and cost (B/C).
High levels of environmental noise may lead to psychological symptoms. The present study has hypothesized that personal listening device (PLD) use and a negative attitude to noise are significantly ...related to anxiety and/or depression.
This is a cross-sectional study conducted on 431 six-grade students (35% male) at the Faculty of Medicine, University of Belgrade. All students completed the questionnaires related to socio-demographic data, noise/music exposure habits, behaviors related to PLD use, tinnitus, headaches, consumption of alcohol, coffee, energy drinks and cigarettes, YANS (Youth Attitude to Noise Scale), CES-D (The Center for Epidemiologic Studies Depression), and GAD-7 (Generalised Anxiety Disorder Assessment - 7 item scale) questionnaires.
There were 396 (91.9%) of investigated students who used PLD, with more frequent use among girls compared to boys (95.0% and 86.8%, respectively; p = 0.002). We found no significant relationship between the frequency of PLD use and depression and anxiety. However, continuous daily PLD use longer than 1 hour was significantly and positively related to depression (p = 0.006). Students with depression had a lower total YANS score compared to those without depression, indicating a more negative attitude toward noise (p = 0.042). Students with no difficulties to concentrate in noise and with a positive attitude to daily noises had about 37% lower chance to suffer from depression and anxiety, respectively.
PLD use is common among medical students and may be associated with their mental health. Longer than 1 hour of continuous daily use of PLD may be positively related to depression. We also found a significant relationship between difficulties concentrating in noisy environments and depression, and between negative attitudes to daily noises and anxiety.
Redirecting waste biomass back into the economy can reduce their burden on the environment. The use of waste biomass for the production of fuels, value-added materials or natural fillers has ...significant economic and environmental benefits. Physico-chemical characterization of waste biomass (plum pomace and fig pomace) was done by proximate, elemental, biochemical analysis, FTIR and SEM analysis. The calorific value of both biomasses can be compared with the calorific value of lignite, which rises their potential use as a solid biofuel. The combustion behavior of biomass was investigated by thermal analysis techniques. Based on thermal degradation experiments performed at four heating rates a kinetic analysis of the biomass decomposition process was accomplished.
The kinetic study was done using Kissinger method, Ozawa method, Flynn-Wall-Ozawa (FWO) method, Starink method, and Kissinger-Akahira-Sunose (KAS) method. The value of activation energy obtained by different kinetic methods was ∼210 kJ/mol for plum pomace and ∼162 kJ/mol for fig pomace. Estimated activation energy values were used to calculate thermodynamic parameters. In addition to the fact that the obtained results can serve as a useful reference for the design of pyrolysis reactors, this research has ecological significance because it solves the problem of solid waste disposal.
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•First detailed report on physicochemical analysis of plum and fig pomace.•The HHV for plum and fig pomace was 17.1 and 16.8 MJ/kg, respectively.•Pyrolysis of plum and fig pomace were studied using non-isothermal TGA.•Detailed report on the kinetics and thermodynamics of plum and fig pomace pyrolysis.•Activation energy for plum pomace is ∼210 kJ/mol and ∼162 kJ/mol for fig pomace.
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