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1.
Environ Res ; 222: 115411, 2023 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-36736753

RESUMEN

The global decline in amphibian populations is a major environmental issue. Chytridiomycosis, Ranaviruses and the red-leg syndrome have been identified in unusual mortality events. However, these infections do not account for all causes of declining amphibian populations. Moreover, several cases of amphibian mortality are difficult to solve without resorting to an interdisciplinary approach. Two cases of unusual mortality in Rana temporaria occurred at two high-mountain ponds (northwest Italy) in April and May 2021. Water and frog samples were analysed to understand the possible causes responsible for the unusual mortalities. Results of the main physicochemical (pH, conductivity, dissolved oxygen, chemical and biochemical oxygen demand) and nutrient (ammonia/ammonium, nitrite, nitrate, total phosphorus) parameters revealed a good condition of the water quality, with the absence of the main cyanotoxins (microcystins/nodularins). However, unseasonably high spring water temperatures were recorded in both ponds (12.73 °C and 14.21 °C for Frog Pond and Selleries Pond, respectively). Frogs (n = 50; snout-vent length: 7.0-9.8 cm; body mass: 85-123 g) collected from Frog Pond mainly presented bumps on the ventral cavity and dermal ulceration associated with the isolation of Carnobacterium maltaromaticum. On the other hand, frogs (n = 5; snout-vent length: 8.0-9.1 cm; body mass: 87-92 g) from Selleries Pond presented petechiae and dermal ulcerations on the rear limbs associated with the isolation of Aeromonas salmonicida and A. sobria. In both mortality events, the interdisciplinary approach revealed an association between frog mortalities and the isolation of bacteria. Isolated bacteria are considered opportunistic pathogens, and the high values of the water temperature has certainly led a stress on the frogs, favouring the spread of bacteria and the death of the frogs. Further studies are needed to assess the pathophysiological effects of the opportunistic bacteria here isolated, clarifying the interactions between emerging pathogens and climate change.


Asunto(s)
Estanques , Ranavirus , Animales , Rana temporaria , Cambio Climático , Anfibios
2.
BMC Vet Res ; 19(1): 21, 2023 Jan 25.
Artículo en Inglés | MEDLINE | ID: mdl-36698186

RESUMEN

BACKGROUND: Hepatitis E Virus (HEV) is recently considered an emerging public health concern. HEV genotypes 1 and 2 are widely distributed and pathogenic only for humans. In contrast, HEV, genotypes 3 and 4 are observed in swine, deer, wild boars and rabbits and can also be transmitted to humans. The presence of HEV in the liver, muscle, faeces, blood, and bile was detected by real-time RT-PCR in 156 pigs belonging to twenty different farms, ranging from 1 to 8 months of age. The phylogenetic analysis was performed on the viral strain present in the positive biological matrix, with the lowest Ct. HEV-IgG and HEV-IgM in the sera were analysed by two different ELISA kits. RESULTS: Twenty-one pigs, i.e., 13.46% of them (21/156, 95% CI: 8.53%-19.84%), tested positive for HEV in at least one biological matrix by real-time RT-PCR, while phylogenetic analysis revealed the presence of HEV subtypes 3f and 3c. Pig serums analysed by ELISA showed an overall prevalence of 26.92% (42/156, 95% CI: 20.14%-34.60%) for HEV-IgG, whereas the 28.95% (33/114, 95% CI: 20.84%-38.19%) of them tested negative resulted positive for the HEV-IgM. CONCLUSIONS: The faeces are the biological matrix with the highest probability of detecting HEV. The best concordance value (Kappa Kohen index) and the highest positive correlation (Phi index) were observed for the correlation between bile and liver, even when the number of positive liver samples was lower than the positive bile samples. This finding may suggest that a higher probability of HEV occurs in the bile, when the virus is present in the liver, during the stages of infection. Finally, the presence of HEV in muscle was observed in 11 pigs, usually used for the preparation of some dishes, typical of the Italian tradition, based on raw or undercooked meat. Therefore, their consumption is a possible source of infection for final consumer.


Asunto(s)
Ciervos , Virus de la Hepatitis E , Hepatitis E , Enfermedades de los Porcinos , Humanos , Porcinos , Animales , Conejos , Virus de la Hepatitis E/genética , Hepatitis E/epidemiología , Hepatitis E/veterinaria , Filogenia , Enfermedades de los Porcinos/epidemiología , Ciervos/genética , Italia/epidemiología , ARN Viral/genética , ARN Viral/análisis , Inmunoglobulina G , Inmunoglobulina M , Sus scrofa/genética
3.
Med Vet Entomol ; 37(3): 472-482, 2023 09.
Artículo en Inglés | MEDLINE | ID: mdl-36715237

RESUMEN

The hippoboscid Lipoptena fortisetosa Maa, 1965 is a hematophagous ectoparasite of cervids that can bite humans. This fly is expanding its geographical range and is of concern for animal and human health since it can potentially harbour harmful microorganisms. This study was aimed at characterizing the bacterial communities of L. fortisetosa in its different life-cycle stages. Pupae and wingless adults were collected from cervids hunted in Tuscan-Emilian Apennines (central Italy) and pooled into groups of 10 by life stage (30 individual pupae; 1420 individual wingless adults). Winged flies were caught by sweep netting and separated into five pools of 10 insects. After DNA extraction, the bacterial content of each pool was analysed using 16 S metabarcoding. Results revealed that the composition and relative abundance of different taxa greatly differed in the three analysed groups. Wingless adults showed a high abundance of Bartonella (33.07%), which is almost absent in winged flies and pupae. Among the detected pathogens, four genera of concern for human health were found: Bartonella, Moraxella, Mycobacterium and Rickettsia. Interestingly reads similar to Bartonella bovis, Moraxella osloensis and Arsenophonus lipopteni Operational Taxonomic Unit (OTUs) were detected. These findings suggest the possible role of L. fortisetosa as a reservoir of pathogenic microorganisms, confirming the need for further investigation to ascertain its vectorial capacity.


Asunto(s)
Bartonella , Ciervos , Dípteros , Rickettsia , Animales , Ciervos/parasitología , Italia , Pupa
4.
J Fish Dis ; 46(7): 731-741, 2023 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-36943008

RESUMEN

Lactococcosis, caused by the Gram-positive bacterium Lactococcus garvieae, is a major concern in rainbow trout (Oncorhynchus mykiss) farms, which are regularly affected by outbreaks especially during the summer/fall months. In these farms, unvaccinated healthy and symptomatic fish can coexist with vaccinated fish. In the present study, innate (leukogram, serum lysozyme activity, peroxidase activity, antiprotease activity, bactericidal activity, total IgM and total proteins), and specific immune parameters (serum antibodies to L. garvieae) were assessed in unvaccinated adult rainbow trout naturally exposed to the pathogen, with or without evidence of clinical signs, or subjected to vaccination. Blood was drawn from all three groups, and blood smears were prepared. Bacteria were found in the blood smears of 70% of the symptomatic fish but not in any of the asymptomatic fish. Symptomatic fish showed lower blood lymphocytes and higher thrombocytes than asymptomatic fish (p ≤ .05). Serum lysozyme and bactericidal activity did not vary substantially among groups; however, serum antiprotease and peroxidase activity were significantly lower in the unvaccinated symptomatic group than in the unvaccinated and vaccinated asymptomatic groups (p ≤ .05). Serum total proteins and total immunoglobulin (IgM) levels in vaccinated asymptomatic rainbow trout were significantly higher than in unvaccinated asymptomatic and symptomatic groups (p ≤ .05). Similarly, vaccinated asymptomatic fish produced more specific IgM against L. garvieae than unvaccinated asymptomatic and symptomatic fish (p ≤ .05). This preliminary study provides basic knowledge on the immunological relationship occurring between the rainbow trout and L. garvieae, potentially predicting health outcomes. The approach we proposed could facilitate infield diagnostics, and several non-specific immunological markers could serve as reliable indicators of the trout's innate ability to fight infection.


Asunto(s)
Enfermedades de los Peces , Infecciones por Bacterias Grampositivas , Oncorhynchus mykiss , Animales , Oncorhynchus mykiss/microbiología , Infecciones por Bacterias Grampositivas/prevención & control , Infecciones por Bacterias Grampositivas/veterinaria , Infecciones por Bacterias Grampositivas/microbiología , Muramidasa , Enfermedades de los Peces/microbiología , Lactococcus , Anticuerpos Antibacterianos , Inmunoglobulina M , Peroxidasas
5.
J Food Sci Technol ; 59(2): 686-692, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-35153311

RESUMEN

Extra virgin olive oil is the highest quality olive oil mainly due to its beneficial constituents and nutritional properties. However, olive oil adulteration is a common fraudulent practice by deliberate mislabelling of less expensive oil categories and admixing expensive olive oils with low oils. To protect consumers from such commercial frauds, an easy and fast method to detect the real composition of oil is needed. For this study we used direct sampling analysis (DSA) coupled with a high-resolution mass spectrometer (AxION2 TOF Perkin Elmer) to analyse the fatty acid composition of three types of edible oil: extra virgin olive oil, refined olive oil and seed oil (EVOO, ROO, and SO respectively) to find a marker that could distinguish between them. Good precision in repeatability and reproducibility (RSD% < 15%) was obtained. The fatty acid ratio between the oleic acid/oleic acid dimer was able to distinguish EVOO from the other two types of oil, while the ratio between linoleic and oleic acid was found to discriminate refined oil from seed oil. The development of an easy, fast and cost-effective method can help to limit commercial frauds, increase the number of controlled samples, and enhance food control along the commercial chain. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13197-021-05063-y.

6.
Fish Shellfish Immunol ; 105: 457-468, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-32673645

RESUMEN

Lactococcosis is one of the main bacterial diseases affecting rainbow trout (Oncorhynchus mykiss), with significant economic and sanitary repercussion. Vaccination and antibiotic treatments are commonly used to prevent and control the infection outbreaks; however, these strategies have some drawbacks including limited coverage, handling costs, induction of antibiotic resistance and chemical residues in the environment. Selective breeding programs represent a promising complementary approach for increasing fish disease resistance in commercial farms and some immunological parameters may be tentatively used as indirect indicators for this purpose. The present study investigated for the first time some innate and adaptive immune responses in two groups of rainbow trout derived from selected lines (susceptible and resistant) showing a different "in field" phenotypical resistance to Yersinia ruckeri, Flavobacterium branchiophilum, F. psychrophilum, and Ichthyophthirius multifiliis, after an immersion-dilution based exposure to Lactococcus garvieae carried out in controlled experimental conditions. Twenty-six resistant and twenty-six susceptible female rainbow trout (mean body weight 80 g, 9 months aged, F5 generation) were obtained from an intensive farm considered L. garvieae free and were exposed to the pathogen. Moreover, 10 resistant and 10 susceptible fish were used as uninfected controls. After 5 days, blood and tissue samples were collected for immunological analyses. A significantly higher serum and mucus lysozyme activity was recorded in resistant rainbow trout compared to susceptible fish (P ≤ 0.05), both before and after exposure to L. garvieae. Similarly, respiratory burst activity of head kidney leukocytes resulted more intense in resistant fish (P ≤ 0.05), suggesting that phagocytes could more quickly activate their microbicidal mechanisms to counteract the bacterial spread. Resistant group displayed also an up-regulation of immunoglobulins M (IgM), major histocompatibility complex II (MHC-II) and interleukin 8 (IL-8) gene expression (P ≤ 0.05) and a significantly higher blood lymphocytes count (P ≤ 0.05), highlighting their potential better ability to trigger the recruitment of defensive cells and the initiation of specific immune processes such as antigen presentation to CD4+ T lymphocytes and IgM synthesis. The results herein presented might be useful for the identification of immunological markers to be used as indirect indicators in rainbow trout selective breeding programs.


Asunto(s)
Resistencia a la Enfermedad/inmunología , Susceptibilidad a Enfermedades/veterinaria , Enfermedades de los Peces/inmunología , Infecciones por Bacterias Grampositivas/veterinaria , Inmunidad Innata , Oncorhynchus mykiss , Inmunidad Adaptativa , Animales , Susceptibilidad a Enfermedades/inmunología , Susceptibilidad a Enfermedades/microbiología , Enfermedades de los Peces/microbiología , Flavobacterium/fisiología , Infecciones por Bacterias Grampositivas/inmunología , Infecciones por Bacterias Grampositivas/microbiología , Hymenostomatida/fisiología , Lactococcus/fisiología , Yersinia ruckeri/fisiología
7.
Vet Res ; 50(1): 97, 2019 Nov 25.
Artículo en Inglés | MEDLINE | ID: mdl-31767033

RESUMEN

Scrapie in goats has been known since 1942, the archetype of prion diseases in which only prion protein (PrP) in misfolded state (PrPSc) acts as infectious agent with fatal consequence. Emergence of bovine spongiform encephalopathy (BSE) with its zoonotic behaviour and detection in goats enhanced fears that its source was located in small ruminants. However, in goats knowledge on prion strain typing is limited. A European-wide study is presented concerning the biochemical phenotypes of the protease resistant fraction of PrPSc (PrPres) in over thirty brain isolates from transmissible spongiform encephalopathy (TSE) affected goats collected in seven countries. Three different scrapie forms were found: classical scrapie (CS), Nor98/atypical scrapie and one case of CH1641 scrapie. In addition, CS was found in two variants-CS-1 and CS-2 (mainly Italy)-which differed in proteolytic resistance of the PrPres N-terminus. Suitable PrPres markers for discriminating CH1641 from BSE (C-type) appeared to be glycoprofile pattern, presence of two triplets instead of one, and structural (in)stability of its core amino acid region. None of the samples exhibited BSE like features. BSE and these four scrapie types, of which CS-2 is new, can be recognized in goats with combinations of a set of nine biochemical parameters.


Asunto(s)
Western Blotting/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Enfermedades de las Cabras/clasificación , Scrapie/clasificación , Animales , Western Blotting/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Europa (Continente) , Enfermedades de las Cabras/diagnóstico , Cabras , Scrapie/diagnóstico
8.
BMC Vet Res ; 15(1): 230, 2019 Jul 06.
Artículo en Inglés | MEDLINE | ID: mdl-31277645

RESUMEN

BACKGROUND: The small ruminant lentiviruses (SRLVs) are a heterogeneous group of viruses that includes caprine arthritis encephalitis virus (CAEV) and Maedi-Visna virus (MVV). SRLVs affect the production and welfare of sheep and goats worldwide. There is currently no effective treatment. Their high mutation rate precludes vaccine development, making innovative control measures necessary. A variant of the chemokine (C-C motif) receptor 5 (CCR5) gene is reportedly involved in resistance to human immunodeficiency (HIV) infection in humans and to SRLV in sheep. The aim of this study was to analyse the genetic structure and variability of the CCR5 gene in goats and to carry out a cross-sectional study to investigate the role of CCR5 genetic variants in controlling susceptibility/resistance to CAEV. RESULTS: The variant g.1059 T located in the promoter region revealed an interesting association with high proviral loads (a 2.8-fold increased risk). A possible explanation could be an alteration of the transcriptional level. Overexpression of the CCR5 receptor on the cell surface may increase virus internalization and proviral load as a consequence. CONCLUSIONS: Our findings could be advantageously used to reduce the susceptibility of goat herds to CAEV by negatively selecting animals carrying the g.1059 T mutation. Eliminating animals predisposed to high proviral loads could also limit the development of clinical signs and the spread of the virus, since these animals are also highly efficient in shedding the virus.


Asunto(s)
Enfermedades de las Cabras/genética , Enfermedades de las Cabras/virología , Infecciones por Lentivirus/veterinaria , Receptores CCR5/genética , Animales , Virus de la Artritis-Encefalitis Caprina , Estudios Transversales , Predisposición Genética a la Enfermedad , Cabras , Infecciones por Lentivirus/genética , Polimorfismo de Nucleótido Simple , Regiones Promotoras Genéticas , Provirus , Análisis de Secuencia de ADN
9.
J Fish Dis ; 42(5): 773-776, 2019 May.
Artículo en Inglés | MEDLINE | ID: mdl-30850994

RESUMEN

This work reports a mortality outbreak, occurred in 2015 and affecting juveniles of European perch (Perca fluviatilis L.) farmed in Italy. Perch rhabdovirus (PRV) was detected by viral isolation and biomolecular investigations. Phylogenetic analysis clustered our isolate into genogroup B, which also includes PRV isolates from Perca fluviatilis identified in France (2004-2009); diagnostic investigations also revealed opportunistic bacteria (Aeromonas hydrophila) and parasites (Chilodonella piscicola). Since, occasionally, PRV has been reported in the natural environment, which is often a source of eggs and broodstock for farms, it could be possible that both similar France and Italian isolate were imported from a same place elsewhere and have a common origin. Improving biosecurity measures (batch control) and disinfection of egg strings with an iodine-based solution helps prevent apparent vertical transmission of PRV.


Asunto(s)
Brotes de Enfermedades/veterinaria , Enfermedades de los Peces/mortalidad , Percas , Infecciones por Rhabdoviridae/veterinaria , Rhabdoviridae/fisiología , Animales , Acuicultura , Enfermedades de los Peces/virología , Italia/epidemiología , Filogenia , Rhabdoviridae/clasificación , Infecciones por Rhabdoviridae/mortalidad , Infecciones por Rhabdoviridae/virología
10.
Acta Vet Hung ; 67(1): 135-139, 2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30922086

RESUMEN

Metagenomic analysis revealed the presence of porcine parvovirus 3 (PPV3) in the pool of the internal organs of a wild boar found dead in Southern Italy. Phylogenetic analysis based on the complete coding sequences showed that the newly detected virus is most closely related to those found also in wild boars in Romania during 2010-2011. Even though the death could not be associated with this virus, PPV3 could have contributed to lowering the host's immunological defences.


Asunto(s)
Metagenómica/métodos , Parvovirus Porcino/genética , Sus scrofa , Enfermedades de los Porcinos/virología , Animales , Genoma Viral , Italia/epidemiología , Filogenia , Porcinos , Enfermedades de los Porcinos/epidemiología
11.
J Hered ; 109(6): 653-662, 2018 08 24.
Artículo en Inglés | MEDLINE | ID: mdl-30010804

RESUMEN

Genealogical relationships among colony members, inbreeding status, and presence of hybrids are crucial data that can assist zoo curators in captive colony management and decision-making on relocation for reproduction. This study employed molecular markers to study a large colony (n = 56) of African Penguin hosted in an Italian biopark. A panel of 15 STRs (single tandem repeats) was selected, and genotype data were analyzed using COLONY software to determine parentage relationships and compare the existing studbook information to a pedigree built from genetic analyses. The existence of extra-pair mating and the presence of hybrids were investigated: discrepancies in kinship relationships emerged following molecular parentage analysis and 10 unknown genetic relationships were revealed. Infidelity of one member of the pair was observed in 6 cases and extra-pair copulation was assessed by genetic analysis in 2 episodes. One member of the colony was found to be a hybrid (S. demersus × S. humboldti); his progeny, derived by extra-pair copulation, was traced. Three other hidden hybrids were discovered and assessed using the identified candidate private alleles. Overall, our results demonstrate that molecular methods to confirm parentage and analyze relatedness among colony members are a valuable tool to complement studbook-based genetic management of African penguin captive populations. Because a variety of behavioral dynamics (e.g., extra-pair mating) can make observations ineffective in some species and because molecular markers outperform studbook in identifying the presence of hybrids, reliance on studbook information alone is not recommended.


Asunto(s)
Hibridación Genética , Spheniscidae/genética , Animales , ADN Mitocondrial , Femenino , Marcadores Genéticos , Técnicas de Genotipaje , Masculino , Análisis de Secuencia de ADN , Análisis para Determinación del Sexo , Spheniscidae/clasificación
12.
J Gen Virol ; 98(7): 1963-1967, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28691895

RESUMEN

The presence of lysine (K) at codon 222 has been associated with resistance to classical scrapie in goats, but few scrapie cases have been identified in 222Q/K animals. To investigate the contribution of the 222K variant to PrPres formation in natural and experimental Q/K scrapie cases, we applied an immunoblotting method based on the use of two different monoclonal antibodies, F99/97.6.1 and SAF84, chosen for their different affinities to 222K and 222Q PrP variants. Our finding that PrPres seems to be formed nearly totally by the 222Q variant provides evidence that the 222K PrP variant confers resistance to conversion to PrPres formation and reinforces the view that this mutation has a protective role against classical scrapie in goats.


Asunto(s)
Enfermedades de las Cabras/metabolismo , Lisina/metabolismo , Proteínas PrPSc/metabolismo , Scrapie/metabolismo , Secuencias de Aminoácidos , Animales , Codón/genética , Codón/metabolismo , Genotipo , Cabras , Lisina/genética , Péptido Hidrolasas/metabolismo , Proteínas PrPSc/química , Proteínas PrPSc/genética , Scrapie/genética
15.
Arch Virol ; 161(2): 471-7, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26597186

RESUMEN

Border disease virus (BDV) is a (+) single-stranded RNA pestivirus affecting mainly sheep and goats worldwide. Genetic typing of BDV has led to the identification of at least seven major genotypes. This study reports the detection of a BDV strain from a goat in northwestern Italy during routine investigations. Sequence analysis revealed mutations in the 5'-UTR of the virus with implications for BDV molecular diagnostics. Moreover, subsequent phylogenetic analysis based on the combined 5'-UTR and Npro/partial C genes, showed divergence from known BDV genotypes, revealing the detection of a novel pestivirus group, for which we propose the name BDV genotype 8.


Asunto(s)
Regiones no Traducidas 5' , Enfermedad de la Frontera/diagnóstico , Enfermedad de la Frontera/virología , Virus de la Enfermedad de la Frontera/clasificación , Virus de la Enfermedad de la Frontera/genética , Genotipo , ARN Viral/genética , Animales , Virus de la Enfermedad de la Frontera/aislamiento & purificación , Análisis por Conglomerados , Cabras , Italia , Datos de Secuencia Molecular , Mutación , Patología Molecular , Filogenia , Análisis de Secuencia de ADN
16.
Ecotoxicology ; 25(2): 431-8, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26597735

RESUMEN

The presence of xenobiotics, such as metals, in ecosystems is concerning due to their durability and they pose a threat to the health and life of organisms. Moreover, mercury can biomagnify in many marine food chains and, therefore, organisms at higher trophic levels can be adversely impacted. Although feathers have been used extensively as a bio-monitoring tool, only a few studies have addressed the effect of both age and sex on metal accumulation. In this study, the concentrations of trace elements were determined in the feathers of all members of a captive colony of African Penguins (Spheniscus demersus) housed in a zoological facility in Italy. Tests were performed by inductively coupled plasma-mass spectrometry to detect aluminum, arsenic, cadmium, cobalt, chromium, copper, iron, manganese, nickel, lead, selenium, tin, vanadium, and zinc. Mercury was detected by a direct mercury analyzer. Sexing was performed by a molecular approach based on analyzing the chromo-helicase-DNA-binding1 gene, located on the sex chromosomes. Sex- and age-related differences were studied in order to investigate the different patterns of metal bioaccumulation between male and female individuals and between adults and juveniles. Juvenile females had significantly higher arsenic levels than males, while selenium levels increased significantly with age in both sexes. Penguins kept in controlled environments-given that diet and habitat are under strict control-represent a unique opportunity to determine if and how metal bioaccumulation is related to sex and age.


Asunto(s)
Animales de Zoológico/metabolismo , Exposición a Riesgos Ambientales , Metales Pesados/metabolismo , Spheniscidae/metabolismo , Oligoelementos/metabolismo , Contaminantes Químicos del Agua/metabolismo , Factores de Edad , Animales , Monitoreo del Ambiente , Plumas/química , Femenino , Italia , Masculino
17.
J Clin Microbiol ; 53(4): 1115-20, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25609728

RESUMEN

Statutory surveillance of bovine spongiform encephalopathy (BSE) indicates that cattle are susceptible to both classical BSE (C-BSE) and atypical forms of BSE. Atypical forms of BSE appear to be sporadic and thus may never be eradicated. A major challenge for prion surveillance is the lack of sufficiently practical and sensitive tests for routine BSE detection and strain discrimination. The real-time quaking-induced conversion (RT-QuIC) test, which is based on prion-seeded fibrillization of recombinant prion protein (rPrPSen), is known to be highly specific and sensitive for the detection of multiple human and animal prion diseases but not BSE. Here, we tested brain tissue from cattle affected by C-BSE and atypical L-type bovine spongiform encephalopathy (L-type BSE or L-BSE) with the RT-QuIC assay and found that both BSE forms can be detected and distinguished using particular rPrPSen substrates. Specifically, L-BSE was detected using multiple rPrPSen substrates, while C-BSE was much more selective. This substrate-based approach suggests a diagnostic strategy for specific, sensitive, and rapid detection and discrimination of at least some BSE forms.


Asunto(s)
Bioensayo/métodos , Encefalopatía Espongiforme Bovina/clasificación , Encefalopatía Espongiforme Bovina/diagnóstico , Priones/análisis , Priones/química , Animales , Química Encefálica , Bovinos , Cricetinae , Encefalopatía Espongiforme Bovina/metabolismo , Humanos , Priones/metabolismo , Proteínas Recombinantes
18.
BMC Genet ; 15: 139, 2014 Dec 14.
Artículo en Inglés | MEDLINE | ID: mdl-25496717

RESUMEN

BACKGROUND: Toll-like receptors play a key role in innate immunity by recognizing pathogens and activating appropriate responses. Pathogens express several signal molecules (pathogen-associated molecular patterns, PAMPs) essential for survival and pathogenicity. Recognition of PAMPs triggers an array of anti-microbial immune responses through the induction of various inflammatory cytokines. The objective of this work was to perform a case-control study to characterize the distribution of polymorphisms in three candidate genes (toll-like receptor 2, toll-like receptor 4, toll-like receptor 9) and to test their role as potential risk factors for tuberculosis infection in water buffalo (Bubalus bubalis). RESULTS: The case-control study included 184 subjects, 59 of which resulted positive to both intradermal TB test and Mycobacterium bovis isolation (cases) and 125 resulted negative to at least three consecutive intradermal TB tests. The statistical analysis indicated that two polymorphisms exhibited significant differences in allelic frequencies between cases and controls. Indeed, the TT genotype at TLR9 2340 C > T locus resulted significantly associated with susceptibility to bovine tuberculosis (P = 0.030, OR = 3.31, 95% CI = 1.05-10.40). One polymorphism resulted significantly associated with resistance to the disease, and included the CC genotype, at the TLR4 672 A > C locus (P = 0.01, OR = 0.26, 95% CI = 0.08-0.80). Haplotype reconstruction of the TLR2 gene revealed one haplotype (CTTACCAGCGGCCAGTCCC) associated with disease resistance (P = 0.04, OR = 0.51, 95% CI = 0.27-0.96), including the allelic variant associated with disease resistance. CONCLUSIONS: The work describes novel mutations in bubaline TLR2, TLR4 and TLR9 genes and presents their association with M. bovis infection. These results will enhance our ability to determine the risk of developing the disease by improving the knowledge of the immune mechanisms involved in host response to mycobacterial infection, and will allow the creation of multiple layers of disease resistance in herds by selective breeding.


Asunto(s)
Polimorfismo de Nucleótido Simple , Receptor Toll-Like 2/genética , Receptor Toll-Like 4/genética , Tuberculosis Bovina/genética , Animales , Búfalos , Estudios de Casos y Controles , Bovinos , Frecuencia de los Genes , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Tuberculosis , Tuberculosis Bovina/inmunología
19.
Dement Geriatr Cogn Disord ; 38(3-4): 245-53, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24969517

RESUMEN

BACKGROUNDS/AIMS: Alzheimer's disease (AD) is one of the main types of dementia affecting about 50-55% of all demented patients. Sleep disturbances in AD patients are associated with the severity of dementia and are often the primary reason for institutionalization. These sleep problems partly resemble the core symptoms of narcolepsy, a sleep disorder caused by a general loss of the neurotransmitter hypocretin. The aim of our study was to investigate whether genetic variants in the hypocretin (HCRT) and in the hypocretin receptors 1 and 2 (HCRTR1, HCRTR2) genes could modify the occurrence and the clinical features of AD and to examine if these possible variants influence the role of the protein in sleep regulation. METHODS: Using a case-control strategy, we genotyped 388 AD patients and 272 controls for 10 SNPs in the HCRT, HCRTR1 and HCRTR2 genes. In order to evaluate which residues belong to the HCRTR2 binding site, we built a molecular model. RESULTS: The genotypic and allelic frequencies of the rs2653349 polymorphism were different (χ(2) = 5.77, p = 0.016; χ(2) = 6.728, p = 0.035) between AD patients and controls. The carriage of the G allele was associated with an increased AD risk (OR 2.53; 95% CI 1.10-5.80). No significant differences were found in the distribution of either genotypic or allelic frequencies between cases and controls in the HCRTR1 polymorphisms rs2271933, rs10914456 and rs4949449 and in the HCRTR2 polymorphism rs3122156. CONCLUSION: Our data support the hypothesis that the HCRTR2 gene is likely to be a risk factor for AD. The increased risk inferred is quite small, but in the context of a multi-factorial disease, the presence of this polymorphism may significantly contribute to influencing the susceptibility for AD by interacting with other unknown genetic or environmental factors in sleep regulation.


Asunto(s)
Enfermedad de Alzheimer/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Neuropéptidos/genética , Receptores de Orexina/genética , Anciano , Anciano de 80 o más Años , Alelos , Estudios de Casos y Controles , Femenino , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Masculino , Orexinas , Polimorfismo de Nucleótido Simple
20.
Animals (Basel) ; 14(12)2024 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-38929341

RESUMEN

Reptiles are usually asymptomatic carriers of Salmonella, with the manifestation of typical clinical signs of acute forms in adult and non-immunocompromised animals being considered exceptions. In the present case, an adult male corn snake (Pantherophis guttatus) was found dead due to septic shock 48 h after consuming a feeder mouse purchased online. The snake's tissue samples and faeces were cultured for bacteria isolation. Microbiological examinations of the snake and mouse livers revealed the presence of Salmonella enterica subsp. enterica serovar Midway. A whole-genome analysis of these two isolates showed a high correlation between them: they belonged to the strain type ST-357 for the classic MLST scheme and to the strain type ST 171322 for the cgMLST scheme. Also, a virulence gene analysis revealed the presence of stdB and STM3026 genes. This report conveys a case of food-borne salmonellosis in a pet snake, transmitted from a feeder mouse, likely responsible for the snake's death due to septic shock. It highlights the relevance of feeder mice as a source of Salmonella infections in snakes and the associated risks to human health.

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