Dynamics of ammonia and pH in meat stored under various conditions

The changes in ammonia content and pH values in beef and pork stored at different temperatures were studied in relation with sensory changes. Samples were taken from the stored meat at a temperature of +3 degrees C in the 0th, 12th, 36th, 60th, 84th, 108th and 132nd hour and at a temperature of +10 degrees C in the 0th, 6th, 24th, 36th, 54th, 72nd and 84th hour. The quantity of ammonia in the samples was determined by means of an ion-selective electrode. The pH value was determined potentiometrically. The kinds of meat (pork, beef) differed in the development of ammonia content. Ammonia content was found to be directly dependent on storage temperature. A similar relation was observed in the development of pH values. The sensory changes of stored meat were parallel to the changes in ammonia levels and pH values. The results can be used for an evaluation of the degree of biochemical changes in the meat and of the fitness of the raw material to the production of tins for long storage. Advantageous points are seen in the rapid and objective determination of ammonia content by the ion-selective electrode and the determination of the pH value by means of the combined glass electrode with the aid of the same apparatus.     

The influence of gibberellic acid, 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine on fruit-set of Clementine mandarin

A study was made of the influences of gibberellic acid (GA₃), 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BA) on leafy inflorescences — and of GA₃ on entire trees — on fruit-set and fruit-weight of Clementine mandarin (Citrus reticulata Blanco).Applying GA₃ to inflorescences of cultivar ‘Fino’ (seedless Clementine) between anthesis and 14 days after anthesis increased fruit-set and weight of the fruits at harvest. No significant differences in effects were observed between the application to leaves, to flowers or to leaves and flowers simultaneously. GA₃ induced the development of parthenocarpic fruits of cultivar ‘Monreal’ (Clementine with seeds) and a fruit-set similar to that obtained with self-pollination. Spraying GA₃ (5–200 mg/l) to entire trees of cultivar ‘Fino’ increased the number of fruits per tree and decreased the average weight of the fruits. However, the commercial yield per tree increased with the concentration of GA₃ employed.The application of 2,4-D and BA to inflorescences of cultivar ‘Fino’ increased fruit-setting and weight of the fruits, although not as efficiently as GA₃, both the concentration and the time of application being more critical.     

Characteristics of the whole cell fatty acid profiles ofPseudomonas corrugate

The fatty acid methyl ester (FAME) profiles of eighty strains ofPseudomonas corrugata from different geographic origins have been studied. Gas chromatographic profiles were obtained. The use of hexane/methyl-tert butyl ether (MTBE) (11) for the extractions improves the yield of hydroxy-FAMEs used to identifyP. corrugata as compared to hexane alone. The analysis of the extracts with hexane/MTBE showed that many strains do not have the characteristic FAMEs ofP. corrugata (3-hydroxydodecenoic acid, 3-hydroxytetradecanoic acid, 3-hydroxyhexadecanoic acid and two unknown additional fatty acids). These differences among strains seemed to be related to bacterial dissociation, associated with changes in morphological aspect of the colonies due to subculture. The comparison of profiles of wrinkled and smooth colonies isolated from ten strains confirmed the differences among those in specific FAMEs. Therefore, the FAME profiles are a useful tool for the identification ofP. corrugata when the bacteria have not been subculturedin vitro for a long time. Multivariate analyses of data showed that four clusters can be observed supporting the heterogeneity of the strains ofP. corrugata.Abbreviations FAME fatty acid methyl ester - MTBE methyl tert-butyl ether     

Antibodies and PMBC from EIAV infected carrier horses recognize gp45 and p26 synthetic peptides

Equine infectious anemia virus (EIAV) is a lentivirus causing a persistent infection in horses characterized by recurrent febrile episodes and high levels of viremia associated with a novel antigenic strain of the virus. The virus contains two envelope glycoproteins, gp90 and gp45, and four internal proteins, p26, p15, p11 and p9. Considering that the most infected horses are able to restrict EIAV replication to very low levels and that gp45 and p26 contain highly conserved epitopes among lentiviruses, it would be necessary to identify those conserved epitopes stimulating cellular and humoral responses. The aims of this study were to determine if the synthetic peptides identified as gp45 (aa 523-547) and p26 (aa 318-346) representing two highly conserved and immunodominant regions of EIA virus are recognized by PBMC and antibodies to EIAV adult mixed-breed naturally infected carrier horses, and if these peptides are able to induce immune responses in mice. Antibodies from 100% of carrier horses, evaluated by ELISA, recognized both peptides; PBMC from 80% of carrier horses, evaluated by lymphoproliferation assay, recognized, at least, one peptide. Furthermore, immunization with 100 microg of each peptide elicited humoral and cellular responses in BALB/c mice, antibodies appeared at 48 or 63 days of immunization with gp45 or p26, respectively. Although the kinetics of gp45- and p26-specific antibody responses were similar, percentage of positivity was higher for gp45. The lymphoproliferation assay, evaluated by BrdU uptake, was higher in mice immunized with gp45 or p26 than in the control group (P<0.05). Based on our findings, we consider that both peptides could be included in an effective vaccine design to induce long-term immunological memory.     

Diagnosis of cystic echinococcosis on sheep farms in the south of Argentina: areas with a control program

In 2000 Guarnera et al. proposed using ELISA in canine faeces collected from the ground to detect dogs infected with Echinococcus granulosus, thus determining sheep farms with active transmission. The objective was to evaluate the prevalence of E. granulosus infection in sheep farms of the Patagonia. Sheep farms were randomly selected in the Provinces of Río Negro, Chubut, Neuquén, Santa Cruz and Tierra del Fuego (areas with control programs) and La Pampa (comparison area). From one to three samples of fecal matter were obtained for each sheep farm, which were processed by means of copro-ELISA test with confirmation of positive samples by copro-Western blot. A total of 1042 samples were obtained from 352 sheep farms, 26 (7.3%) proving positive. Of these 5 (6.3%) were from La Pampa, 9 (13.8%) from Neuquén, 4 (4.7%) from Río Negro, 2 (2.9%) from Chubut, 1 (5.9%) from Santa Cruz and 5 (13.9%) from Tierra del Fuego. The identification of parasitized dogs is an essential activity upon which rests the strategy of control and surveillance. Arecoline tests or coproantigen test with fecal matter obtained directly from the dog contribute information on individual prevalence, while the use of coproantigens detected in ground-collected samples transfers the dog unit of observation to units of greater epidemiological value. In the present experience, the technique employed seems promising for its application in systems of epidemiological surveillance of cystic echinococcosis and in drawing a baseline on which to measure the progress of control programs in the Argentine Patagonia in subsequent years.     

Stereological study of the external urethral sphincter in the female urethra of the lamb: a new model for studies on urinary continence

Some of the morphometrical features and functional properties of the lamb external urethral sphincter (EUS) were determined by means of ATPase histochemistry. In this study, eight female urethras of 2- to 3-month-old lambs were used. The samples were studied by using an unbiased stereological design and were statistically evaluated with a Kolmogorov-Smirnov test. Additionally, the mechanical activity of the different fibre types was analysed by measuring the isometric changes in tension induced by electrical field stimulation (EFS) of intrinsic nerves. The lamb EUS has an average fibre density of 1142.37 fibres/mm(2) (SD = 279.33). Our data showed the predominance of fast-twitch fibres or type II fibres, with a proportion of 30% (type I) and 70% (type II) in all cases. These percentages remained the same along the length of the EUS. However, a clear tendency to increase the number of both fibre types in the mid-EUS was observed. Interestingly, type I and II fibres are intermingled throughout the whole lamb EUS. Dynamic parameters of the contraction induced in lamb EUS preparations by EFS of intramural nerves, were intermediate between fast- and slow-twitch muscles. Furthermore, contractile activity showed no decrease in amplitude upon repetitive stimulation, suggesting the presence of a mixture of fibre types, in which fast but fatigue-resistant fibres (probably type IIA) seem to predominate. Our results indicate that lambs could be suggested as a new biological model to study those pathological processes, which involve the sphincteric regulation of the EUS.     

Development of a new PCR assay to identify Brucella abortus biovars 5, 6 and 9 and the new subgroup 3b of biovar 3

One hundred twenty-nine Brucella field strains isolated from cattle in Cantabria, Spain, from March 1999 to February 2003, were analysed by using the AMOS-ERY PCR assay and by Southern blot hybridisation with a probe from insertion sequence IS711. Most of the field isolates produced only the ery band in the AMOS-ERY assay and showed a hybridisation pattern identical to that exhibited by reference strains of biovars 5, 6 and 9 of Brucella abortus, but different from strain Tulya, belonging to biovar 3 of B. abortus. However, typing of these strains by standard methods demonstrated that they belonged to biovar 3 of B. abortus. These results indicated that B. abortus biovar 3 was not genetically homogeneous and at least could be divided in two. In one class, that we called biovar 3a, would be the Tulya strain, while the local field strains would belong to biovar 3b. Cloning and nucleotide sequencing of a DNA fragment containing an IS711 copy exclusive of the B. abortus field strains from biovar 3b and reference strains from biovars 5, 6 and 9, revealed the existence of a 5.4 kb deletion close to an IS711 copy. Based on these data, we designed a new primer, which together with the IS711 AMOS primer produced a PCR fragment of 1.7 kb only from the isolates of biovars 3b, 5, 6 and 9 of B. abortus. No amplification products were produced with these primers from strains of the rest of species and biovars of Brucella and from bacteria phylogenetically close to Brucella analysed in this work. Addition of this primer to the AMOS-ERY PCR primer cocktail allows the positive distinction of B. abortus biovars 3b, 5, 6 and 9 from the rest of Brucella species and biovars.     

Resistance to <Emphasis Type="Italic">Penicillium</Emphasis><Emphasis Type="Italic">hirsutum</Emphasis> Dierckx in garlic accessions

Among the factors affecting the quality and yield of garlic production, blue mold caused by -- Penicillium spp. -- is responsible for economical losses in many countries. Allicin, present in garlic bulbs, has been suggested as having antifungal activity against some Penicillium species. This study was conducted to evaluate the response of garlic accessions against Penicillium hirsutum infection and to compare this response with bulb allicin content. Twelve garlic accessions were inoculated with P. hirsutum, and assayed in greenhouse and growth chamber experiments. Plant growth parameters and the fungal production of conidia were evaluated. Significant differences were found among the accessions. Accessions Castaño and Morado were most resistant whereas AR-I-125 and Fuego were always severely affected by the disease. A low correlation was found (r = 0.17) between allicin content and tolerance, indicating that allicin is not the main factor involved in the resistance against P. hirsutum.     

Detection and identification of equine Theileria and Babesia species by reverse line blotting: epidemiological survey and phylogenetic analysis

Specific oligonucleotide probes were designed to develop a new and highly sensitive reverse line blot assay to detect and identify simultaneously different Theileria and Babesia species in horses. The amplified hypervariable V4 region of the 18S rRNA gene was hybridised against different generic and species-specific probes. The survey was conducted over 243 samples of equine blood divided into three different groups: group 1, 24 horses presented as possible clinical piroplasmosis; group 2, 181 clinically healthy free-ranging horses exposed to ticks; group 3, 38 riding horses with unrelated pathologies and low or no contact with ticks. The study demonstrated a high piroplasm prevalence in the first two groups of animals. Two Theileria genotypes sharing 96.8% similarity between their 18S rRNA gene sequences and two Babesia genotypes sharing 97.4% similarity, were identified. The biologic meaning of such genotypes is discussed in terms of their phylogenetic relationships and potential pathogenicity.     

Failure to establish infection with Tetratrichomonas sp. in the reproductive tracts of heifers and bulls

Experimental infection of the reproductive tracts of heifers and bulls with Tetratrichomonas sp. isolated from preputial smegma of virgin bulls was attempted. Nine heifers and four bulls were challenged by inoculation of 7 x 10(6) Tetratrichomonas sp. into the vaginal lumen and preputial cavity, respectively. Vaginal mucus and preputial smegma samples were collected and cultured for Tetratrichomonas sp. Heifers were slaughtered in groups of three at 2, 9 and 21 days after inoculation. Two heifers and two bulls infected with Tritrichomonas foetus and two uninfected heifers were used as controls for the model infection. Tetratrichomonas sp. were only isolated in vaginal mucus of 7/9 inoculated heifers at 6h post-inoculation, and genital secretions taken at slaughter time from vagina, uterus and oviduct were cultural negative. Bulls challenged with Tetratrichomonas sp. remained cultural negative. Since Tetratrichomonas sp. survived only a few hours in the female genitalia and did not survive in the male genitalia after experimental challenge, Tetratrichomonas sp. did not colonize the genital tract. These were likely trichomonads from the digestive tract. Collection of clean samples without fecal contamination from the reproductive tract is proposed as a measure to avoid Tetratrichomonas sp. transitory genital infection.