Microbiological and molecular characterization of Corynebacterium diphtheriae isolated in Algeria between 1992 and 2015.

The objectives of this study were to undertake the microbiological and molecular characterization of Corynebacterium diphtheriae isolates collected in Algeria during epidemic and post-epidemic periods between 1992 and 2015. Microbiological characterization includes the determination of biotype and toxigenicity status using phenotypic and genotypic methods. Antimicrobial susceptibility was determined by the E-test method. Molecular characterization was performed by multi-locus sequence typing. In total, there were 157 cases of C. diphtheriae isolates, 127 in patients with respiratory diphtheria and 30 with ozena. Isolates with a mitis biotype were predominant (122 out of 157; 77.7%) followed by belfanti (28 out of 157; 17.8%) and gravis biotype (seven out of 157; 4.5%). Toxigenic isolates were predominant in the period 1992-2006 (74 out of 134) whereas in the period 2007-2015, only non-toxigenic isolates circulated (23 out of 23). All 157 isolates were susceptible to erythromycin, gentamicin, vancomycin and cotrimoxazole. Reduced susceptibility to penicillin G, cefotaxime, tetracycline and chloramphenicol was detected in 90 (57.3%), 88 (56.1%), 112 (71.3%) and 90 (57.3%) isolates, respectively. Multi-locus sequence typing analysis indicates that sequence type 116 (ST-116) was the most frequent, with 65 out of 100 isolates analysed, in particular during the epidemic period 1992-1999 (57 out of 65 isolates). In the post-epidemic period, 2000-2015, 13 different sequence types were isolated. All belfanti isolates (ten out of 100 isolates) belonged to closely related sequence types grouped in a phylogenetically distinct eBurst group and were collected exclusively in ozena cases. In conclusion, the epidemic period was associated with ST-116 while the post-epidemic period was characterized by more diversity. Belfanti isolates are grouped in a phylogenetically distinct clonal complex.

Detection of urease-negative Bordetella bronchiseptica from the field.

Four urease-negative Bordetella bronchiseptica isolates originating from pigs were examined by phenotypic and molecular methods. The phenotypic properties of the isolates were in harmony with the data of the literature, except for the lack of urease activity in conventional tube test, API 20 NE and Diatabs™ assays. Using genotypic methods, the urease-negative isolates did not differ from the urease-positive reference strain. They were positive in species-specific and ureC PCR, and all strains showed uniform bands in PCR-RFLP studies of flaA genes. The reason for the lack of urease activity, a characteristic considered species specific for B. bronchiseptica, needs to be studied further. The finding underlines the significance of genotyping when the phenotypic identification of B. bronchiseptica seems questionable.

Prevalence of clinical signs of disease in Danish finisher pigs.

Between December 1999 and February 2001, two visits, eight weeks apart, were made to 90 herds of Danish finisher pigs. The prevalence of clinical signs was recorded by three veterinary technicians from the Danish Bacon and Meat Council according to a standardised procedure; they had been trained and their observations were monitored and validated before and during the study. A total of 154,347 finisher pigs were examined and 22,136 clinical signs were recorded. Vices accounted for 43 per cent of the signs. The highest mean prevalence was observed for ear necrosis (4.44 per cent), followed by respiratory signs (2.17 per cent), lameness (1.92 per cent), other skin diseases (1.73 per cent), tail bites (1.26 per cent), umbilical hernia (0.78 per cent), flank bites (0.52 per cent), diarrhoea (0.27 per cent), respiratory distress (0.12 per cent), atrophic rhinitis (0.10 per cent), recumbency (0.09 per cent) and central nervous disease (0.05 per cent). The prevalence of atrophic rhinitis was higher in conventional herds than in specific pathogen-free herds. The prevalence of clinical signs of atrophic rhinitis was higher among finishers weighing 51 to 75 kg than among finishers weighing up to 50 kg, and the prevalence of respiratory signs was higher among finishers weighing 51 to 75 kg then among finishers weighing 76 to 100 kg.

Differentiation of Pasteurella multocida isolates from cases of atrophic rhinitis in pigs from Zimbabwe by RAPD and ribotyping.

Atrophic rhinitis in pigs is rarely reported in Southern Africa. To determine the relationship between Pasteurella multocida clones from clinical cases of atrophic rhinitis, twenty-one strains were characterised by selected phenotypic and genotypic methods. Biochemical analysis classified 18 strains as P. multocida subspecies multocida, whilst the remainder were grouped into separate unassigned biotypes. Capsular groups A (16/21) and D (l/21) were found among the isolates by PCR. Four ribotype patterns were obtained following HpaII ribotyping, whilst random amplification of polymorphic DNA (RAPD) revealed three main clusters. However, subclusters were also noted for each RAPD cluster. Our results indicate that RAPD offers a better discrimination of strains than ribotyping and that none of the phenotypic characters were directly related to the genotypic clusters.

Persistent rhinitis - allergic or nonallergic?

Although rhinitis has been classified as being either allergic, noninfectious, or "other forms" (nonallergic noninfectious), these categories lack strict classification criteria and often overlap. The term "nonallergic noninfectious rhinitis" is commonly applied to a diagnosis of any nasal condition, in which the symptoms are similar to those seen in allergic rhinitis but an allergic aetiology has been excluded. This group comprises several subgroups with ill-defined pathomechanisms, and includes idiopathic rhinitis, irritative-toxic (occupational) rhinitis, hormonal rhinitis, drug-induced rhinitis, and other forms (non-allergic rhinitis with eosinophilia syndrome [NARES], rhinitis due to physical and chemical factors, food-induced rhinitis, emotion-induced rhinitis, atrophic rhinitis). Unlike allergic rhinitis, there are no specific diagnostic tests and diagnosis is primarily based on a history of reaction to specific irritant-toxic triggering agents (either general or occupational), drugs, infections, and hormonal status, coupled with exclusion of allergic rhinitis and other forms of non-allergic rhinitis by skin prick testing. Accordingly, from a clinical standpoint NARES, irritative-toxic, hormonal, drug-induced and idiopathic rhinitis are possibly the least difficult forms of nonallergic rhinitis to diagnose.

Frecuencia de rinitis atrófica y grado de lesión de los cornetes nasales de cerdos en Yucatán, México / Frequency of atrophic rinitis and the degree of lesions in pig nasal turbinates from Yucatan, Mexico

Introducción.- El complejo respiratorio de los cerdos es uno de los principales síndromes que afectan a la industria porcina, debido al efecto negativo que tiene sobre los costos de producción por concepto de mortalidad, tratamientos, disminución en la ganancia diaria de peso y aumento en la conversión alimenticia. El objetivo de este estudio fue determinar la frecuencia de las lesiones macroscópicas en cornetes de cerdos finalizados obtenidos en el mercado municipal No. 32 de Mérida, Yucatán, México. Materiales y Métodos.- Se inspeccionaron 101 hocicos de cerdos de engorda finalizados adquiridos en el mercado municipal No. 32 de Mérida, Yucatán, México, de abril a junio de 1997. Con un rinómetro se determinó el grado de rinitis. A los casos positivos a rinitis atrófica progresiva (RAP) se les realizó un análisis histopatológico y bacteriológico. Resultados.- Cincuenta y cinco (54.5 por ciento) hocicos presentaron RAP. Treinta y tres (60.0 por ciento) cornetes nasales presentaron grado 2, 17 (30.9 por ciento) cornetes nasales con grado 3 y 5 (9.1 por ciento) cornetes nasales con grado 4. Se observó 1 (1.8 por ciento) caso con cuerpos de inclusión y correspondió al grado 4 de RAP. Se aislaron Pasteurella multocida en 2 (3.6 por ciento) casos positivos a RAP y correspondieron a los grados 2 y 4 y Bordetella bronchiseptica en 3 (5.5 por ciento) muestras y correspondieron a los grados 2, 3 y 4 de RAP. Discusión.- Las diferencias encontradas entre la RAP de este estudio y las reportadas por los diferentes autores, probablemente se debieron a la técnica utilizada. La baja frecuencia de aislamientos probablemente se debió a que las muestras fueron tomaron de cerdos sacrificados en el rastro, lo cual pudo favorecer la contaminación de las muestras. La RAP probablemente se asocia a los agentes etiológicos B bronchiseptica, P multocida y Citomegalovirus. En el estado de Yucatán, México, este es el primer reporte de un caso de rinitis atrófica con cuerpos de inclusión. Esto permite demostrar que la RAP es un padecimiento que no necesariamente guarda relación con un solo agente etiológico.

Development of a typing system for epidemiological studies of porcine toxin-producing Pasteurella multocida ssp. multocida in Denmark.

The aim of the present study was to evaluate capsular-typing, plasmid-profiling, phage-typing and ribotyping for epidemiological studies of toxin-producing Pasteurella multocida ssp. multocida in Denmark. The evaluation of methods was based on 68 strains from nasal swabs and 14 strains from pneumonic lungs. Strains from lungs were all of capsular Type A, whereas strains from nasal swabs were of both capsular Types A and D. Only 9% of the strains contained plasmids, which could not be associated with antibiotic resistance. Phage-typing divided 61% of strains into 10 groups, while 39% were non-typable. CfoI ribotyping divided strains into four groups of which one type contained 94% of isolates. HindIII ribotyping divided strains into 18 types. A total of 18 strains from The Netherlands, UK and USA were subjected to HindIII ribotyping, resulting in 13 types of which six were identical to ribotypes of Danish strains. Phage-typing of isolates from an outbreak of atrophic rhinitis involving six herds in 1985 showed the existence of an epidemic strain. This type was recognised in the herd suspected of being the source of the infections and in four of the five infected herds. These findings were supported by HindIII ribotyping, as 85% of isolates from all herds were assigned to one ribotype. In conclusion, HindIII ribotyping seems to represent a useful tool for epidemiological studies of toxigenic P. multocida ssp. multocida.

Associations between behavioural indicators of "stress" in weaner pigs and respiratory lesions at slaughter.

The aim of the study was to investigate associations on the individual-pig level between eight behavioural indicators of 'stress' (defined in a very general sense), in seven to eight-week old piglets four weeks after weaning and the occurrence of atrophic rhinitis (AR) and chronic pleuritis (CP) at slaughter. The study was designed as a cohort study, including 512 pigs from two commercial farms. With one exception Actinobacillus pleuropneumoniae serotype 2 (App2) infection was not associated with any of the behaviour patterns observed. Using random-effects logistic regression, weak statistical associations, positive as well as negative, were found between the performance of several behaviour patterns and the two pathological lesions. This suggests the existence of biological associations between oral behaviour patterns in young pigs and the health status later in life. However, the results were not consistent between specific oral behaviour patterns or between the two herds indicating that the associations found are either due to chance or reflect the existence of herd-specific confounders. Therefore, results based on studies within only one herd should not be generalized to other herds without care. Behavioural observations on seven- to eight-week-old piglets on the basis of the present investigation cannot be recommended for use as early predictors of AR or CP at slaughter. Further studies are needed in order to elucidate causal relations between behaviour and health.

Demonstration that Australian Pasteurella multocida isolates from sporadic outbreaks of porcine pneumonia are non-toxigenic (toxA-) and display heterogeneous DNA restriction endonuclease profiles compared with toxigenic isolates from herds with progressive atrophic rhinitis.

Capsular types A and D of Pasteurella multocida cause economic losses in swine because of their association with progressive atrophic rhinitis (PAR) and enzootic pneumonia. There have been no studies comparing whole-cell DNA profiles of isolates associated with these two porcine respiratory diseases. Twenty-two isolates of P. multocida from diseased pigs in different geographic localities within Australia were characterised genotypically by restriction endonuclease analysis (REA) with the enzyme CfoI. Seven of 12 P. multocida isolates from nasal swabs from pigs in herds where PAR was either present or suspected displayed a capsular type D phenotype. These were shown to possess the toxA gene by polymerase chain reaction (PCR) and Southern hybridisation, and further substantiated by production of cytotoxin in vitro. The CfoI profile of one of these seven isolates, which was from the initial outbreak of PAR in Australia (in Western Australia, WA), was identical with profiles of all six other toxigenic isolates from sporadic episodes in New South Wales (NSW). The evidence suggests that the strain involved in the initial outbreak was responsible for the spread of PAR to the eastern states of Australia. Another 10 isolates, representing both capsular types A and D, were isolated exclusively from porcine lung lesions after sporadic outbreaks of enzootic pneumonia in NSW and WA. CfoI restriction endonuclease profiles of these isolates revealed considerable genomic heterogeneity. Furthermore, none of these possessed the toxA gene. This suggests that P. multocida strains with the toxA gene do not have a competitive survival advantage in the lower respiratory tract or that toxin production does not play a role in the pathology of pneumonic lesions, or both. REA with polyacrylamide gel electrophoresis and silver staining was found to be a practical and discriminatory tool for epidemiological tracing of P. multocida outbreaks associated with PAR or pneumonia in pigs.

[A preliminary study on etiology of atrophic rhinitis in Zunyi].

200 cases (households with atrophic rhinitis (AR) diagnosed from outpatients and mobile medical team as the AR group were studied. Another 200 cases (households) served as the control group, who were residents of countryside and members of a factory in Zunyi, and they have lived in Zunyi for more than 20 years. The subjects between the two groups were similar in age and sex. Each of these two groups was further divided into two groups, the opentype stove group (100 cases) and the closetype stove group (100 cases). The heritability (h2, 59.4 +/- 0.032%) was estimated by using the method of Falconer liability threshold. The concentration of SO2 in living environment was monitored with Parafuchsin colorimetry. The daily average SO2 concentrations have been drawn for comparison. The result showed that the SO2 concentration of living environment in AR group was significantly higher than that in control group (P < 0.01). We believe that the contraction and development of AR were determined by the combination of inheritance with environment. This fact suggests that AR may be transmitted in a multigenic and multifacterial modes.

An epidemiologic and economic study of respiratory diseases in two conventional Danish swine herds. I: Prevalence of respiratory lesions at slaughter and their effects on growth.

A total of 578 slaughter pigs from 2 Danish conventional farrow-to-finish operations (Herds A and B) were followed from an age of 14 days to slaughter. Pigs were weighted at 3 weeks intervals and at slaughter and an extended post mortem examination of the plucks was done. Comparison of growth rates in pigs with and without specific types of lesions by the t-test and those with multiple lesions with regression models demonstrated that Mycoplasma-like pneumonia, complicated pneumonia, anterio-ventral pleuritis, fissures and atrophic rhinitis significantly reduced mean daily gain and increased the time required to reach slaughter weight. The total impact of the lesions in Herd A was an estimated reduction in mean daily gain of 27 grams and a 2 day increase in the interval from 14 days of age until slaughter (MDG14). Decreases in MDG14 in Herd B were more substantial, 98 grams and 16.7 days. Reductions in mean daily gains during the interval from the fourth weighing until slaughter were 31 grams in Herd A and 137 grams in Herd B. Chronic dorso-caudal and parietal pleuritis, without other lesions present, had no significant adverse effects on growth rates in either herd. Interactions between lesions did not significantly alter the estimates. The R2 values obtained for the regression models showed that the presence, absence or extent of lesions at slaughter explained only 13-27% of the variations in growth rates in the 2 herds.

Relationship to growth performance of pneumonia and atrophic rhinitis lesions detected in pigs at slaughter among four seasons.

A commercial swine herd was selected for study, because pigs at slaughter repeatedly had lung lesions consistent with enzootic pneumonia and had snout lesions typical of atrophic rhinitis. Pigs born during various seasons of the year were allotted to 4 investigations and were evaluated from birth to slaughter. Individual lungs and snouts were identified and collected at the slaughter plant and later examined for gross lesions of bronchopneumonia and atrophic rhinitis, respectively. Each lesion was scored, and the following comparisons were made within investigations: prevalence and mean scores for lung lesions; prevalence and mean grades for snout lesions; correlations between lung lesion scores and growth indicators; correlations between snout lesion grades and growth indicators; and correlations between lung lesion scores and snout grade scores. Included in the growth indicators were average daily gain during the growing phase, average daily gain during the finishing phase, average daily gain during growing and finishing phases, and days to attain 104.5 kg of body weight. Prevalence of lung or snout lesions, mean values for lung lesion scores, mean values for snout lesion grades, and mean values for the various growth indicators were tested for statistical differences among the 4 investigations. Prevalence of lung lesions was highest (96%) for winter-slaughtered and lowest (81%) for autumn-slaughtered pigs. Mean scores for lung lesions were 7% (summer), 5% (autumn), 9% (winter), and 16% (spring). Prevalence of snout lesions was highest (85%) for spring-slaughtered pigs and lowest (42%) for autumn-slaughtered pigs.(ABSTRACT TRUNCATED AT 250 WORDS)

Restriction endonuclease analysis of porcine Pasteurella multocida isolates from Quebec.

We have used restriction endonuclease analysis (REA) of genomic DNA to classify porcine Pasteurella multocida isolates with similar capsular and somatic serotypes, and to monitor the distribution of isolates from 12 different herds in Quebec. Within herds, P. multocida isolates of similar capsular and somatic serotypes showed similar REA fingerprints. Between herds, some isolates had similar REA fingerprints. However, differences in REA enabled subtyping of many P. multocida isolates with the same antigen types. Our data indicate that REA would enable accurate epidemiological typing of P. multocida in conjunction with classical capsular and somatic typing.

Prevalência de rinite atrófica e de pneumonia em granjas associadas a sistemas de integraçäo de suínos, no estado de Santa Catarina / Prevalence of atrophic rhinits and pneumonia in farms associated to swuine integration systems in the State of Santa Catarina

No inverno de 1985 examinaram-se suínos terminados abatidos em frigoríficos, oriundos de 133 granjas de suínos associadas a cinco sistemas de integraçäo em Santa Catarina, para avaliar a prevalência de rinite atrófica e de pneumonias. As granjas foram escolhidas de forma aleatória, em média 25 por integraçäo. De cada granja foi examinado um lote, em média, de 27 animais de diferentes raças e ambos os sexos, com idade variando de cinco meses e meio a seis meses e meio. Dos 3.607 cornetos nasais examinados, 52,6% näo apresentavam lesöes macroscópicas, 31,4% estavam ligeiramente afetados (grau 1), 11,9% apresentavam atrofia definida (grau 2) e 4,9% atrofia grave (grau 3). Dos 3.588 pulmöes examinados, 44,7% näo apresentavam lesöes, 44,9% , mostravam hepatizaçäo em 0 < X < 11% do pulmäo (grau 2), 7,6% com 11 < X < 21% (grau 3) e 2,9% com X > 21% (grau 4). A rinite atrófica e a pneumonia foram diagnosticadas em 100% das granjas. Concluiu-se que a rinite atrófica e a pneumonia estäo amplamente disseminadas nos rebanhos produtores de suínos terminados associados aos sistemas de integraçäo no Estado de Santa Catarina

Ascariasis, respiratory diseases and production indices in selected Prince Edward Island swine herds.

The levels of production, ascarid burden and respiratory disease were measured on 15 purposively selected swine herds, and the relationships between the various measures of ascarid burden were examined. On each farm 30 randomly selected pigs were weighed and rectal fecal samples were collected at approximately 11, 15, 19 and 22 weeks of age, and at slaughter. Fecal ascarid-egg counts and duration of infection were combined to calculate a composite measure of ascarid burden called "lifetime burden". At the abattoir the carcass weight and levels of anteroventral pneumonia, atrophic rhinitis, and liver lesions were recorded for each hog. The number of ascarids in the small intestines were counted. Study hogs were marketed at an average of 189 +/- 22 days. The average dressed carcass weight was 77.0 +/- 5.9 kg and the mean average daily gain was 0.519 +/- 0.071 kg/day. The percent of hogs with ascariasis varied widely among farms, no matter what measure of ascariasis was used; the percent with intestinal ascarids at slaughter ranged from 0% to 96%, the percent that shed ascarid eggs during their lifetime ranged from 0% to 100%, and the range for hogs with liver lesions ranged from 27% to 100%. Of the hogs slaughtered, 82% had milk spot lesions, 32% shed ascarid eggs during their lifetime and 35% had intestinal ascarids. The latter had an average of 12 intestinal ascarids. Anteroventral pneumonia occurred in 55% of the slaughtered hogs and 9% had atrophic rhinitis scores of five. The percent of hogs per farm with pneumonia ranged from 17% to 96%. The percent of hogs per farm with atrophic rhinitis scores of five ranged from 0% to 57%.

Development and epidemiological applications of a bacteriophage typing system for typing Pasteurella multocida.

A bacteriophage typing system was developed for typing toxigenic and nontoxigenic strains of Pasteurella multocida. A phage set of 24 phages with different lytic spectra was isolated after mitomycin treatment of P. multocida strains, isolated mainly from pigs from herds with atrophic rhinitis. On a test set of 97 different strains isolated from pigs, these 24 phages were able to type 87% of the strains. The 97 test strains could be subdivided into 31 different types by reaction with the 24 phages. The reproducibility after subculture and storage of the strains was good (95%). Phage typing of 217 toxigenic P. multocida field isolates from 37 pig herds predominately with clinically atrophic rhinitis resulted in 18 different phage types and an overall typability of 68%. Of 24 herds from which more than three isolates of toxigenic P. multocida were obtained, a single phage type was demonstrated in 5 herds, while in 9 herds a single phage type represented at least half of the isolates. The phage types in the remaining 10 herds revealed no dominating phage type. The phage typing system described appears to be a valuable epidemiological tool for studying the spread of P. multocida.

Toxigenic type D Pasteurella multocida in New South Wales pig herds--prevalence and factors associated with infection.

Between March and July 1987, a study was undertaken to determine the prevalence of and factors associated with toxigenic type D Pasteurella multocida infection in New South Wales pig herds. Toxigenic type D P. multocida was isolated from the nasal cavities of pigs in one (2%) of 50 randomly selected herds. Toxigenic isolates were also recovered from 2 (8%) of a separate group of 25 herds that had purchased pigs from a known infected piggery in South Australia (herd SA). Snout abnormalities were present in 9.4%, 3.2% and 1.8% of grower pigs in the 3 affected herds. Isolation of toxigenic P. multocida was significantly associated (p less than 0.0001) with the occurrence of clinically affected pigs in the herd. Purchase of at least 5 pigs from herd SA was associated with an elevated risk (p less than 0.05) of isolation of toxigenic P. multocida.