Comparison of Variable-Number Tandem-Repeat Markers typing and IS1245 Restriction Fragment Length Polymorphism fingerprinting of Mycobacterium avium subsp. hominissuis from human and porcine origins

Background

Animal mycobacterioses are regarded as a potential zoonotic risk and cause economical losses world wide. M. avium subsp. hominissuis is a slow-growing subspecies found in mycobacterial infected humans and pigs and therefore rapid and discriminatory typing methods are needed for epidemiological studies. The genetic similarity of M. avium subsp. hominissuis from human and porcine origins using two different typing methods have not been studied earlier. The objective of this study was to compare the IS1245 RFLP pattern and MIRU-VNTR typing to study the genetic relatedness of M. avium strains isolated from slaughter pigs and humans in Finland with regard to public health aspects.

Methods

A novel PCR-based genotyping method, variable number tandem repeat (VNTR) typing of eight mycobacterial interspersed repetitive units (MIRUs), was evaluated for its ability to characterize Finnish Mycobacterium avium subsp. hominissuis strains isolated from pigs (n = 16) and humans (n = 13) and the results were compared with those obtained by the conventional IS1245 RFLP method.

Results

The MIRU-VNTR results showed a discriminatory index (DI) of 0,92 and the IS1245 RFLP resulted in DI 0,98. The combined DI for both methods was 0,98. The MIRU-VNTR test has the advantages of being simple, reproducible, non-subjective, which makes it suitable for large-scale screening of M. avium strains.

Conclusions

Both typing methods demonstrated a high degree of similarity between the strains of human and porcine origin. The parallel application of the methods adds epidemiological value to the comparison of the strains and their origins. The present approach and results support the hypothesis that there is a common source of M. avium subsp. hominissuis infection for pigs and humans or alternatively one species may be the infective source to the other.     

Atrial-based pacing for sinus node dysfunction in dogs: initial results

Background

An important consideration for the treatment of sick sinus syndrome (SSS) lies in the function of the atrioventricular (AV) node because most patients with SSS retain the ability to conduct atrial impulses.

Hypothesis/Objectives

This retrospective study examined the feasibility of atrial pacing (AAI) in dogs with sinus node dysfunction (SND).

Animals

Sixteen dogs with SND and AAI pacing were identified.

Methods

Retrospective review of medical records.

Results

Follow‐up time ranged from 45 to 1,227 days (mean: 292 days). Only 1 dog developed AV block 3 days postoperatively. Complete lead dislodgment occurred in 3/16 dogs 1, 19, and 27 days postoperatively. Lead perforation into the pericardial space occurred in 2/16 dogs. Rising thresholds for pacing with possible lead microdislodgment or fibrosis were suspected in another 3/16 dogs 57, 192, and 1,016 days after implantation. None of these dogs had complete loss of capture but all required higher thresholds for pacing.

Conclusions and Clinical Importance

Based on this small group of dogs, clinically important AV block does not appear to occur in the long‐term for dogs with SND. Risks of lead perforation, complete dislodgment, and rising thresholds for pacing, possibly because of microdislodgment, may be related to the initial skill level of the operator or the leads that were used. Use of leads with reduced torque at the lead tip, higher flexibility, increased lead‐tip surface of contact with the endocardium or, more likely, use of alternate locations for pacing in the small right atrium of dogs with SND may decrease the frequency of these complications.     

Enantiospecific ketoprofen concentrations in plasma after oral and intramuscular administration in growing pigs

Background

Ketoprofen is a non-steroidal anti-inflammatory drug which has been widely used for domestic animals. Orally administered racemic ketoprofen has been reported to be absorbed well in pigs, and bioavailability was almost complete. The objectives of this study were to analyze R- and S-ketoprofen concentrations in plasma after oral (PO) and intra muscular (IM) routes of administration, and to assess the relative bioavailability of racemic ketoprofen for both enantiomers between those routes of administration in growing pigs.

Methods

Eleven pigs received racemic ketoprofen at dose rates of 4 mg/kg PO and 3 mg/kg IM in a randomized, crossover design with a 6-day washout period. Enantiomers were separated on a chiral column and their concentrations were determined by liquid chromatography-tandem mass spectrometry. Pharmacokinetic parameters were calculated and relative bioavailability (F_rel) was determined for S and R –ketoprofen.

Results

S-ketoprofen was the predominant enantiomer in pig plasma after administration of the racemic mixture via both routes. The mean (± SD) maximum S-ketoprofen concentration in plasma (7.42 mg/L ± 2.35 in PO and 7.32 mg/L ± 0.75 in IM) was more than twice as high as that of R-ketoprofen (2.55 mg/L ± 0.99 in PO and 3.23 mg/L ± 0.70 in IM), and the terminal half-life was three times longer for S-ketoprofen (3.40 h ± 0.91 in PO and 2.89 h ± 0.85 in IM) than R-ketoprofen (1.1 h ± 0.90 in PO and 0.75 h ± 0.48 in IM). The mean (± SD) relative bioavailability (PO compared to IM) was 83 ± 20% and 63 ± 23% for S-ketoprofen and R-ketoprofen, respectively.

Conclusions

Although some minor differences were detected in the ketoprofen enantiomer concentrations in plasma after PO and IM administration, they are probably not relevant in clinical use. Thus, the pharmacological effects of racemic ketoprofen should be comparable after intramuscular and oral routes of administration in growing pigs.     

Population structure of Mycobacterium bovis isolates from cattle in Mexico

The molecular fingerprints of 878 isolates of Mycobacterium bovis collected from cattle between 2009 and 2010 in different regions of Mexico were used in this study. One hundred and ninety-four spoligotypes were observed in total with a high degree of heterogeneity. Sixty-four percent of the isolates grouped into just nine spoligotypes, and 27% fell into only two spoligotypes: SB0673 and SB0669; 149 were orphan spoligotypes. The two predominant spoligotypes were found in almost all states in Mexico, especially in central Mexico, where there is a high concentration of dairy cattle; however, some spoligotypes were closely associated with restricted geographical areas. The hypothetical evolutionary relationship among spoligotypes was estimated using the spoligoforest program in the spolTools webpage. Four trees with connected components and nine unconnected nodes were found. The biggest tree had SB0140 strain as a root, suggesting this as the oldest strain in the tree. However, the relationship of this spoligotype with SB0673 and SB0669 was weak. The discriminatory power of spoligotyping for this M. bovis sample of isolates was 0.94, and the recent transmission index (RTI) 0.83, suggesting a high rate of recent transmission of some strains of M. bovis in the population. This parameter indicates that new measures are required to stop the dissemination of tuberculosis in cattle.     

Bacterial communities and predicted nitrogen metabolism of heterotrophic‐ and probiotic‐based biofilms used for super‐intensive indoor shrimp culture

Biofilm‐based aquaculture systems constitute a promising alternative for intensive shrimp rearing. Microorganisms forming biofilms can recycle the nitrogen compounds the production units improving the water quality while using zero or limited water exchange. This study aimed to compare the taxonomic profiles and the predicted functions related to the transformation of nitrogenous compounds between a heterotrophic‐ (HtB) and a probiotic biofilm (PrB), and the effect of these on the water quality and the productive response of cultured shrimp. Libraries of the 16S‐rRNA gene (V3‐V4 region) were prepared and sequenced to be used as a taxonomic biomarker. Analysis of metagenomic datasets revelated that genera Halomonas, Planctomycetes and Rhodopirellula were the most abundant genera in HtB; meanwhile, Bacillus, Halobacillus and Flavobacterium dominated in PrB. Regarding nitrogen metabolism, the proportion of genes encoding enzymes catalyzing the six pathways shaping the nitrogen metabolism showed differences between biofilms, which could also explain the difference in water quality between treatments. Concerning the productive response of shrimp, no significant differences were detected except for survival, which was higher in PrB. Finally, the results suggest that biofilms harbour functions for nitrogen metabolism, including dissimilatory nitrate reduction, assimilatory nitrate reduction, denitrification, nitrification, nitrogen fixation and anammox; however, the balance of these functional capabilities seems to be relevant to maintain water quality.     

Management of hydrops amnion in a mare resulting in birth of a live foal

CASE DESCRIPTION: A 19-year-old Thoroughbred mare was evaluated at 265 days of gestation with a markedly distended abdomen and edema of the ventral portion of the abdomen. CLINICAL FINDINGS: The uterus was distended over the pelvic rim, making transrectal palpation of the fetus impossible. Transabdominal ultrasonography revealed excessive amounts of fetal fluid. Results of analysis of fluid obtained via amnio- and allantocentesis confirmed that the amniotic cavity was large. TREATMENT AND OUTCOME: The mare was monitored for signs of weakness of the prepubic tendon and abdominal wall. The fetus and placenta were monitored for signs of stress and pending abortion. Flunixin meglumine and altrenogest were administered to the mare. Parturition was attended and occurred at 321 days' gestation. Postpartum complications in the mare included hypovolemic shock and cardiac arrhythmias. Both conditions were treated, and the mare recovered. The foal was considered small, had bilateral angular limb deformities, and was unable to nurse. The foal was given plasma for failure of passive transfer of immunity. Ten months later, the foal underwent procedures to correct limb deformities. CONCLUSIONS AND CLINICAL RELEVANCE: Hydrops conditions are rare in horses, with hydrops allantois occurring more frequently than hydrops amnion; reportedly result in fetal or neonatal death; and may result in death of or injury to the mare. Close monitoring of maternal and fetal health in combination with supportive treatment of the mare can result in the safe progression of a hydrops pregnancy and the birth of a live foal.     

Fruit development in Capsicum annuum: changes in capsaicin, lignin, free phenolics, and peroxidase patterns

Pepper fruits, of Capsicum annuum cv. Padron, undergo changes in content of capsaicinoids, lignin, and free phenolics during the maturation process. Although capsaicinoids increase with development, the maximal levels of free phenolics and lignin are observed during the early stages of development. A decrease of peroxidase activity was observed during maturation, and this was related with a decrease in other physiological parameters studied, namely chlorophylls and pH. Subcellular fractionation studies reveal that most peroxidase activity is localized in the soluble fraction throughout development. The changes in the peroxidase activity were accompanied by changes in the different isoenzymes. Acidic isoenzymes increased whereas the basic isoenzymes decreased over the same period, and the changes in these isoenzymes were related with capsaicin metabolism.     

Low-power switching of phase-change materials with carbon nanotube electrodes

Phase-change materials (PCMs) are promising candidates for nonvolatile data storage and reconfigurable electronics, but high programming currents have presented a challenge to realize low-power operation. We controlled PCM bits with single-wall and small-diameter multi-wall carbon nanotubes. This configuration achieves programming currents of 0.5 microampere (set) and 5 microamperes (reset), two orders of magnitude lower than present state-of-the-art devices. Pulsed measurements enable memory switching with very low energy consumption. Analysis of over 100 devices finds that the programming voltage and energy are highly scalable and could be below 1 volt and single femtojoules per bit, respectively.     

Multiorgan engraftment of human somatic cells in swine foetuses after intra-blastocyst transplantation

Adult human stem cells, mainly from hematopoietic lineage, have been injected into developing pre-immune animal foetuses, and xenogenic engraftment of liver and other organs has been reported. We isolated a rare cell population from adult human liver, fat and skin. Colonies with few cells became visible as early as 2-3 days, and a fully formed colony took 10-14 days to form. These colonies were named as liver-derived cell lines (LDCs), fat-derived cell lines (FDCs) and skin-derived cell lines (SDCs). All these cells express few pluripotency markers like Klf4, c-myc and Sox2. Pig blastocysts were injected with LDCs, FDCs and SDCs and transferred to recipient pigs. We achieved an overall pregnancy rate of 71.4% at day 35. The foetuses were analysed for human cell chimerism in liver, kidney and heart both by RT-PCR and real-time PCR using primers specific to human and pig mitochondrial DNA. The percentage of foetuses showing chimerism was 17.4% (4/23), 12.5% (2/16) and 11.1% (1/9) for LDCs, FDCs and SDCs, respectively. Of these, 42.9% (three out of seven) showed chimerism in liver and 71.4% (five out of seven) showed kidney chimerism. However, we did not detect any chimerism in the heart. The level of chimerism varied and was in the range of one human cell per one hundred thousand to one million pig cells.