Protein insertion into the mitochondrial inner membrane by a twin-pore translocase

The mitochondrial inner membrane imports numerous proteins that span it multiple times using the membrane potential Deltapsi as the only external energy source. We purified the protein insertion complex (TIM22 complex), a twin-pore translocase that mediated the insertion of precursor proteins in a three-step process. After the precursor is tethered to the translocase without losing energy from the Deltapsi, two energy-requiring steps were needed. First, Deltapsi acted on the precursor protein and promoted its docking in the translocase complex. Then, Deltapsi and an internal signal peptide together induced rapid gating transitions in one pore and closing of the other pore and drove membrane insertion to completion. Thus, protein insertion was driven by the coordinated action of a twin-pore complex in two voltage-dependent steps.     

First report of the use of mucosal swabs of the palatine tonsillar crypt for detection of Mycoplasma bovis in naturally infected calves

ABSTRACT

Aims: To compare detection by real-time PCR of DNA from Mycoplasma bovis on mucosal swabs taken from the palatine tonsillar crypt and the mainstem bronchi of clinically asymptomatic calves after slaughter.

Methods: We compared the sensitivity of mucosal swabs taken from two sites: the palatine tonsillar crypt and the mainstem bronchi. Paired samples were taken post-mortem at slaughter from 55 clinically well calves from an infected herd and were tested by real-time PCR for the presence of M. bovis-specific DNA.

Results: Mycoplasma bovis DNA was detected in 51 palatine tonsillar crypt swabs (92.7 (95% CI?=?82.4–98.0)%) and seven mainstem bronchial swabs (12.7 (95% CI?=?5.3–24.5)%). All seven calves with positive mainstem bronchial swabs also had positive palatine tonsillar crypt swabs.

Conclusions: When compared to mucosal swabs of the mainstem bronchi, mucosal swabs of the palatine tonsillar crypt were seven times more sensitive for the post-mortem detection of M. bovis DNA. The viability of detected M. bovis was not assessed, because any cattle carrying viable or non-viable M. bovis DNA were determined to be a potential risk to eradication. Palatine tonsillar crypt mucosa may be a useful anatomical site for real-time PCR detection of M. bovis DNA in naturally infected calves. More work is needed to define the persistence and viability of M. bovis at this anatomical site.

Clinical relevance: The results of this study helped form the basis of surveillance tools used in M. bovis control and eradication efforts. Familiarity with these results may help veterinarians better communicate with their clients about the science behind the eradication efforts.     

An Infection in Chickens with a Strain of Mycoplasma gallisepticum of Low Virulence

A slow-spreading infection due to a strain of Mycoplasma gallisepticum was detected serologically in a flock which remained free of clinical symptoms. The strain was shown to be tylosin resistant. Serological studies indicated that vertical transmission occurred.     

Studies of Escherichia coli Infection in Chickens

The pathogenesis of infection with Escherichia coli was studied in chickens using live O78:K80 cells and a heat-labile chick lethal toxin. The results obtained were compared with those observed in field outbreaks. The common histological findings of subepicardial edema and congestion, focal necrosis in the spleen and focal necrosis, congestion, edema and accumulation of fibrin in the liver support an active role for chick lethal toxin in the pathogenesis of E. coli disease.     

Classical natural ovine scrapie prions detected in practical volumes of blood by lamb and transgenic mouse bioassays

Scrapie is diagnosed antemortem in sheep by detecting misfolded isoforms of prion protein (PrP^Sc) in lymphoid follicles of the rectal mucosa and nictitating membranes. Assay sensitivity is limited if (a) the biopsy is collected early during disease development, (b) an insufficient number of follicles is collected, or (c) peripheral accumulation of PrP^Sc is reduced or delayed. A blood test would be convenient for mass live animal scrapie testing. Currently approved techniques, however, have their own detection limits. Novel detection methods may soon offer a non-animal-based, rapid platform with detection sensitivities that rival the prion bioassay. In anticipation, we sought to determine if diseased animals could be routinely identified with a bioassay using B lymphocytes isolated from blood sample volumes commonly collected for diagnostic purposes in small ruminants. Scrapie transmission was detected in five of six recipient lambs intravenously transfused with B lymphocytes isolated from 5~10 mL of blood from a naturally scrapie-infected sheep. Additionally, scrapie transmission was observed in 18 ovinized transgenic Tg338 mice intracerebrally inoculated with B lymphocytes isolated from 5~10 mL of blood from two naturally scrapie-infected sheep. Based on our findings, we anticipate that these blood sample volumes should be of diagnostic value.     

An overview of dropped hock syndrome cases in New Zealand cattle

ABSTRACT

Case history: Data were collected from 47 outbreaks of dropped hock syndrome (DHS) that were reported by veterinarians in New Zealand to the Ministry for Primary Industries between October 2012 and August 2017. There were 181 affected dairy cows from 44 farms. Of those with records, all 86 were aged between 2–3 years-old, and 4/94 (4%) were Friesian, 56/94 (60%) were Jersey and 33/94 (35%) were Jersey/Friesian cross. Of the 47 outbreaks, 45 (96%) occurred during winter and 37 (79%) in the South Island.

Clinical findings: Of 151 cases with records, hindlimb weakness (117 cows), shortened gait (112 cows) and dropped hocks (106 cows) were most commonly reported, with 110 cases being bilaterally affected. The level of diagnostic work-up and the data recorded by veterinarians for each outbreak were highly variable. Creatine kinase and aspartate aminotransferase activities were reported for 22 cases and were within normal ranges for cows with mild disease but increased in cows with severe disease. Concentrations of Cu in serum and liver were below normal for 13/22 and 9/10 cows, respectively, from six outbreaks. Of 41 cows with records for clinical outcomes, 12 had complete resolution, 18 had partial resolution, and 11 had no resolution.

Pathological findings: Post-mortem data were available from 26 cases. Muscle necrosis and haemorrhage of the origin of the gastrocnemius muscle were the key gross findings. In 14 cows with records for muscle histopathology, myodegeneration and recent haemorrhage were observed, and connective tissue pathology was reported to predate muscular pathology in seven cases.

Diagnosis: DHS appeared to be a degenerative rather than inflammatory condition primarily affecting the connective tissue of the hind limb, especially at the dense collagen interfaces of the gastrocnemius. Although no evidence of neuropathy was found, this cannot definitively be ruled out as a potential cause.

Clinical relevance: A more standardised and systematic approach to investigating cases and recording case data is required to make robust inferences about the aetiology, risk factors, and treatment interventions for DHS.     

Studies on Mycoplasma mycoides subsp. mycoides (LC) in lambs and calves.

Six cesarean-derived lambs were inoculated either with 4.5 X 10(4), 4.5 X 10(6) or 4.5 X 10(8) Mycoplasma mycoides subsp. mycoides intratracheally. One animal receiving the intermediate dose died four days post-inoculation, the two receiving the high dose died six days postinoculation, while one receiving the low dose died eight days postinoculation. The two surviving lambs were challenged on day 20 postinoculation with 1 X 10(8) organisms subcutaneously and 2 X 10(9) organisms intravenously. One animal died eight days following this challenge while the other survived and was killed. Six conventionally reared lambs challenged with 90 to 8500 organisms by intranasal and intraocular instillation failed to become infected. Three conventionally reared calves were each inoculated with 1 X 10(8) organisms by each of intratracheal, subcutaneous and intravenous routes. They were killed 20 days post-inoculation without having shown any clinical signs.     

Studies on the Chick-lethal Toxin of Escherichia coli

A toxin which is lethal for two week old chicks has been recovered from strains of Escherichia coli O78:K80 of bovine and avian origin and from avian isolates of serogroups O2, O45 and O109. The toxin is heat-labile, antigenic, high in protein, inactivated by pronase, trypsin, amylase, and pancreatic lipase. The toxin may be precipitated by ammonium sulfate or TCA treatment from the supernatant obtained by repeated centrifugation of sonicated cells. Considerable purification has been obtained by column chromatography using Sepharose 6B.     

Direct measurement of long-range third-order coherence in Bose-Einstein condensates

A major advance in understanding the behavior of light was to describe the coherence of a light source by using correlation functions that define the spatio-temporal relationship between pairs and larger groups of photons. Correlations are also a fundamental property of matter. We performed simultaneous measurement of the second- and third-order correlation functions for atoms. Atom bunching in the arrival time for pairs and triplets of thermal atoms just above the Bose-Einstein condensation (BEC) temperature was observed. At lower temperatures, we demonstrated conclusively the long-range coherence of the BEC for correlation functions to third order, which supports the prediction that like coherent light, a BEC possesses long-range coherence to all orders.     

Evaluation of factors that affect embryonic loss in dairy cattle

OBJECTIVE: To identify potential risk factors for embryonic loss before 35 to 42 days of gestation in dairy cattle. DESIGN: Prospective observational study. ANIMALS: 381 cows. PROCEDURE: Body condition score was determined at the time of artificial insemination (AI; day 0) and on days 20, 23, and 27 and between days 35 and 41; serum progesterone concentration was measured on days 0; 20 or 21; and 23, 24, or 25. Cows were excluded from analyses if day 0 serum progesterone concentration was > or = 1.0 ng/mL and classified as pregnant on day 23 if serum progesterone was > 1.5 ng/mL on day 20 or 21 and day 23, 24, or 25. Cows were examined via transrectal ultrasonography on day 27 or 28 and rectally palpated for pregnancy on days 35 to 41. RESULTS: 39% of cows that were pregnant on day 23 lost their embryo by day 27, and 18% of cows that were pregnant on day 27 or 28 were not pregnant on days 35 to 41. Breeding a pregnant cow posed the greatest risk for embryonic loss at both time periods. Mean serum progesterone concentrations on day 21 or 22 and day 23, 24, or 25 were lowest for cows that lost an embryo between days 24 and 28. Cows with a linear somatic cell count score > 4.5 before AI were twice as likely to lose the embryo by 35 to 41 days, compared with cows with a score < 4.5. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that embryonic loss could be reduced by more accurate estrus detection, reducing mastitis, and strategies to improve progesterone concentration after breeding.