New hosts for the mite Ornithonyssus bursa in Argentina.

The mite Ornithonyssus bursa (Berlese) (Mesostigmata: Macronyssidae) is considered a poultry pest causing important infestations in chickens and it is considered a potential vector of arbovirus. Despite being considered a common parasite in wild birds, there is scarce published information about its potential hosts and effects on them. Here we present new bird hosts for O. bursa, assess the presence of Alphavirus, Flavivirus and Bunyavirus in mites from three host species, and discuss its potential impact on wild bird populations. We found O. bursa infecting five raptor and six passerine wild bird species. For nine of these species, this is the first record of infection by O. bursa. Although all analysed mites were negative for the examined arboviruses, the small sample size of mites does not allow further conclusions at the present moment. Because of the general nature of this ectoparasite, its presence in migratory long dispersal and endangered bird species, and the seropositivity for arboviruses in some of the species studied here, we consider it critical to assess the role of O. bursa and other ectoparasites as vectors and reservoirs of pathogens and as potential deleterious agents in wild bird populations.

Understanding the relationship between Mycobacterium bovis spoligotypes from cattle in Latin American countries.

Spoligotyping is the most frequently used method for genotyping isolates of Mycobacterium bovis worldwide. In the current work, we compared spoligotypes from 1684 M. bovis isolates from Argentina (816), Brazil (412), Chile (66), Mexico (274) and Venezuela (116), obtained from cattle, humans, pigs, wild boars, farmed deer, goats, buffaloes, cats, and wild animals. A total of 269 different spoligotypes were found: 142 (8.4%) isolates presented orphan spoligotypes, whereas 1542 (91.6%) formed 113 different clusters. In cattle, SB0140 was the most representative spoligotype with 355 (24.6%) isolates, followed by SB0121 with 149 (10.3%) isolates. Clustering of spoligotypes ranged from 95.2% in Argentina to 85.3% in Mexico. Orphan spoligotypes were also variable, ranging from 23.7% in Mexico to 4.1% in Brazil. A large proportion of spoligotypes were common to the neighboring countries Argentina, Brazil and Chile. In conclusion, despite the diversity of spoligotypes found in the five countries studied, there are major patterns that predominate in these neighboring countries. These clusters may reflect a long-lasting active transmission of bovine tuberculosis or common historical origins of infection.

Characterization of Bordetella bronchiseptica strains using phenotypic and genotypic markers.

Thirty-five strains of Bordetella bronchiseptica, recovered primarily from pigs, rabbits, dogs, cats and humans, were characterized by phenotypic and genotypic markers. Biochemical typing only showed variation in the ability to reduce nitrate to nitrite. OMP profiles from virulent strains showed variations in the region of 85-95kDa, which lead us to describe five OMP-types alpha, beta, gamma, delta and epsilon. Genotypic markers included the presence of IS1001, and polymorphisms in the flagellin gene (flaA) and pertussis toxin (PT) promoter region. The IS1001 was detected in 16 isolates (2 from humans and 10 from pigs) but was absent in rabbit isolates. The restriction profiles of the flaA gene allowed us to differentiate the strains into types A-C. The PT types were characterized by an RFLP assay and could be typed through patterns III-V. There was no apparent association between the flaA or PT types and the origin of the isolates. Eleven groups of isolates were identified on the basis of specific combinations of the analyzed markers. The combination of phenotypic and genotypic tests used could be useful in characterizing isolates and differentiating between certain clonal types of B. bronchiseptica.

Descriptive study of human and bovine tuberculosis in Querétaro, México.

A 5-year retrospective study (1992-1996) to look at the situation of human tuberculosis was conducted in Querétaro, México. Also, a 6-month study to determine the frequency of gross lesions in dairy cattle at slaughter, and a short experiment to evaluate the effect of sodium borate in the survival of M. bovis in lesions were carried out. The number of cases were 114 in 1992, 211 in 1995, and 174 in 1996. Possible risk factors were: overcrowding, under-nutrition, previous cases of TB in the family, concurrent Diabetes mellitus, poor personal hygiene, smoking, and alcohol abuse. Eighty percent of the cases were pulmonary. The number of cases increase with age, from 5% in patients 10-year old or younger to 42% in patients 50-year old or older. Seventy-two percent were cured, and only 6% die. Persistent coughing was by far the most observed clinical symptom. From 112 acid-fast negative samples, 8.9% were positive by culture. From 1,201 carcasses revised at slaughter, 17% presented TB-gross lesions. Ninety-six percent were localized lesions involving frequently a single organ, mostly retropharyngeal, mediastinal, mesenteric and mandibular lymph. From 102 lesions, 95% were TB-compatible, and 79% were positive to isolation of M. bovis. Most affected animals were female > 2 years old. It was observed that keeping lesions in a 6% sodium borate solution does not affect the diagnosis of M. bovis by culture after 150 days.

[Enterovirus determination in cases with a diagnosis of the Guillain-Barré syndrome by using the acid-concentration technic]. / Determinación de enterovirus en casos con diagnóstico de síndrome de Guillain-Barré mediante la utilización de la técnica de concentración ácida.

The purpose of this work was to detect enteroviruses in feces by an acid concentration technique (ACT). Fifty-eight samples from children less than 5 years age with diagnosis of Guillain-Barré syndrome (GBS) were analyzed using the routine technique and ACT. Nine positive samples with the routine technique were used as controls. Nine control samples and 22 additional (31 cases) non-polio enteroviruses were isolated and identified with the ACT (53%). Thus, 38% more isolates were obtained by ACT. Isolation was more successful in the RD cellular line (59%) than in Hep-2c (41%). In most cases most titers (71%) obtained were low. ACT improved the detection of enteroviruses but because it is very expensive and laborious, it should be used in the case of laboratories that analyze multiple samples, for special cases such as with autopsy cases and when results are compatible to poliovirus using the routine technique and only in samples obtained during the first 15 days of symptomatology.

Influencia de diferentes derivados lácteos sobre el crecimiento de Listeria monocytogenes en medios selectivos / Influence of different dairy products on the growth kinetic of Listeria monocytogenes

Se estudió la influencia del añadido de crema de leche y leche parcialmente descremada sobre la cinética de crecimiento de Listeria monocytogenes en caldos de enriquecimiento para listerias, conteniendo diferentes concentraciones de acriflavina (15 y 7,5 mg/l). El crecimiento de Listeria monocytogenes en los caldos de enriquecimiento sufrió un retardo atribuible, al menos parcialmente, a la presencia de acriflavina. El añadido de crema de leche o leche parcialmente descremada al caldo de enriquecimiento que contiene 7,5 mg/l de acriflavina produjo un alargamiento de la fase de adaptación, pero las cosechas máximas alcanzadas a las 48 h no mostraron diferencias significativas. En presencia de 15 mg/l de acriflavina, se observó una pérdida inicial de la viabilidad de los cultivos, que fue potenciada por el agregado de crema de leche o leche parcialmente descremada al caldo de enriquecimiento. Además, la leche descremada produjo una disminución de la velocidad máxima de crecimiento que impidió alcanzar la cosecha máxima dentro de las 48 h. Los resultados obtenidos sugieren la necesidad de validar la metodología de recuperación de L. monocytogenes para cada producto, pues la eficiencia de recuperación podría ser afectada por la composición del mismo, sobre todo cuando la carga microbiana es baja

Influencia de diferentes derivados lácteos sobre el crecimiento de Listeria monocytogenes en medios selectivos / Influence of different dairy products on the growth kinetic of Listeria monocytogenes

Se estudió la influencia del añadido de crema de leche y leche parcialmente descremada sobre la cinética de crecimiento de Listeria monocytogenes en caldos de enriquecimiento para listerias, conteniendo diferentes concentraciones de acriflavina (15 y 7,5 mg/l). El crecimiento de Listeria monocytogenes en los caldos de enriquecimiento sufrió un retardo atribuible, al menos parcialmente, a la presencia de acriflavina. El añadido de crema de leche o leche parcialmente descremada al caldo de enriquecimiento que contiene 7,5 mg/l de acriflavina produjo un alargamiento de la fase de adaptación, pero las cosechas máximas alcanzadas a las 48 h no mostraron diferencias significativas. En presencia de 15 mg/l de acriflavina, se observó una pérdida inicial de la viabilidad de los cultivos, que fue potenciada por el agregado de crema de leche o leche parcialmente descremada al caldo de enriquecimiento. Además, la leche descremada produjo una disminución de la velocidad máxima de crecimiento que impidió alcanzar la cosecha máxima dentro de las 48 h. Los resultados obtenidos sugieren la necesidad de validar la metodología de recuperación de L. monocytogenes para cada producto, pues la eficiencia de recuperación podría ser afectada por la composición del mismo, sobre todo cuando la carga microbiana es baja (AU)

[Effect of different dairy products on the growth of Listeria monocytogenes in selective media]. / Influencia de diferentes derivados lácteos sobre el crecimiento de Listeria monocytogenes en medios selectivos.

We have studied the influence of the incorporation of milk cream and skim milk on the growth kinetic of Listeria monocytogenes in listeria enrichment broth with 15 mg/l or 7.5 mg/l of acriflavine. Acriflavine was responsible, at least partially, for delayed growth of Listeria monocytogenes in the enrichment broths. A longer lag phase of the growth was produced by the addition of milk cream or skim milk to the enrichment broth containing 7.5 mg/l of acriflavine. However, the maximum population obtained at 48 h did not show significant differences. In the presence of 15 mg/l of acriflavine, we observed a decrease of the viable counts during the early phase of the growth cycle, which was enhanced by the addition of milk cream or skim milk. Moreover, the maximum growth rate was reduced by the addition of skim milk and maximum population was not reached at 48 h. These results suggest the need to validate the methodology of recuperation of Listeria monocytogenes from each dairy product, since its efficiency may be affected by product composition, specially when the sample biocharge is low.

[Viability of Bacillus subtilis auxotrophs in the absence of their essential metabolites]. / Viabilidad de Bacillus subtilis auxotrofos en ausencia de sus metabolitos esenciales.

The effect of tryptophan and uracil starvation on the viability of the transformant B. subtilis BSA 170 trp- ura- and its parent strains Bacillus subtilis PB 168 trp -C and Bacillus subtilis PB 3308 ura- was examined. These studies were performed at the conditions for competence development, during 16 hours. Our results showed that B. subtilis BSA 170 was resistant to tryptophan-less death during all the assay and was also resistant to uracil-less death during three hours. After this time, viability measurements revealed less colony forming units per milliliter, and decrease of the culture absorbances. The uracil-less death required the presence of tryptophan suggesting that protein synthesis is needed. The parental strains exhibit similar behavior. Bacillus subtilis PB 168 was resistant to tryptophan-less death and B. subtilis PB 3308 showed decrease of the viability after uracil starvation comparable to that of the transformant strain.

Viabilidad de Bacillus subtilis auxotrofos en ausencia de sus metabolitos esenciales. / [Viability of Bacillus subtilis auxotrophs in the absence of their essential metabolites]

The effect of tryptophan and uracil starvation on the viability of the transformant B. subtilis BSA 170 trp- ura- and its parent strains Bacillus subtilis PB 168 trp -C and Bacillus subtilis PB 3308 ura- was examined. These studies were performed at the conditions for competence development, during 16 hours. Our results showed that B. subtilis BSA 170 was resistant to tryptophan-less death during all the assay and was also resistant to uracil-less death during three hours. After this time, viability measurements revealed less colony forming units per milliliter, and decrease of the culture absorbances. The uracil-less death required the presence of tryptophan suggesting that protein synthesis is needed. The parental strains exhibit similar behavior. Bacillus subtilis PB 168 was resistant to tryptophan-less death and B. subtilis PB 3308 showed decrease of the viability after uracil starvation comparable to that of the transformant strain.