First report of a phytoplasma associated with Spondias purpurea (Jocote de Corona) in El Salvador

A new disease of the fruit tree Jocote de Corona (Spondias purpurea L.) was observed in El Salvador, Central America. The symptoms included small chlorotic leaves, highly proliferating shoots, and shortened internodes. Absence of sweet pulp in fruits made them inedible, causing considerable yield losses for farmers. The disease etiology was investigated using polymerase chain reaction with phytoplasma-specific primers, sequencing, and phylogenetic analysis. We obtained no amplification products from symptomless plants, whereas all tests were positive from plants with symptoms. Phylogenetic analysis indicated that this phytoplasma clustered in the 16S rIII group, the type member of which is X-disease phytoplasma. This is the first report of a phytoplasma associated with Jocote de Corona disease in El Salvador and Central America. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AJ888471     

Ovine sperm DNA oxidation quantification using an 8‐OHdG immunodetection assay

Our aim was to optimize 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG) immunodetection in order to detect DNA damage caused by oxidative stress that may not be detected by other DNA integrity analysis techniques, especially due to the high compaction of DNA in ruminants. Semen samples from 6 rams were cryopreserved. After thawing, samples were subjected to the DNA oxidation quantification using an 8‐OHdG immunodetection assay by flow cytometry. We have evaluated two different incubation times (30 min vs. overnight) at 4°C of the primary antibody (monoclonal anti‐8‐OHdG antibody). We have also compared the results of this technique with the sperm chromatin structure assay (SCSA^®). The analysis revealed that there were no significant differences (p > .05) between different incubation times. However, overnight incubation seems to cause more non‐specific binding of the secondary antibody. Significant differences (p < .05) between subjects and oxidation controls (8 M H₂O₂/800 μM FeSO₄?7H₂O) were evident. We can conclude that the 8‐OHdG immunodetection assay for DNA oxidation quantification of ram sperm can be performed subjecting sperm samples to a very high oxidative treatment.     

Physical response of dogs supplemented with fish oil during a treadmill training programme

The rise in popularity of dog sports competitions has led to the evaluation of improvements in dog physical performance. The potential benefit of dietary supplementation with fish oil (FO) on the physical performance of human beings and horses has been reported. However, such effect has not been studied in dogs. We therefore evaluated the effect of FO dietary supplementation on heart rate (HR), rectal temperature (RT) and thigh circumference (TC) in dogs during aerobic treadmill training, and further determined HR response and blood lactate (BL) concentration during an incremental exercise test. Using a cross‐over design, eight male dogs were randomly assigned to two groups and received a standard balanced commercial diet (control, CG, n = 7) and the same diet supplemented with 54 mg FO/kg metabolic weight per day (FOG, n = 8). All dogs had 30‐min treadmill sessions at 8 km/hr and 7.5% slope twice a week for 12 weeks. Assessment of HR and RT was performed before and immediately after each session; HR was also assessed 5 min after the end of each session. Thigh circumference was evaluated before each session. All dogs performed an incremental exercise test on the treadmill at 0, 6 and 12 weeks to evaluate HR response and BL concentration. Data were analysed using the mixed procedure (SAS 9.4). In FOG, pre‐HR (?4.9%) and post‐HR (?2.4%) values and post‐RT (?0.3%) values were lower during treadmill training, whereas TC (+2.2%) values were higher as compared with CG (p < 0.01). Through the incremental exercise test, mean HR (week 6, ?5.3%; week 12, ?6.0%) values in FOG were lower than in CG (p < 0.05). In conclusion, FO supplementation slightly improved the physiological response of dogs to exercise during training.     

Lack of growth of heat-shocked Staphylococcus aureus surviving cells in common media and enterotoxin release.

The ratio of the different types of S. aureus surviving cells immediately after sub-lethal heat treatment (52 degrees C for 15 min) was studied. A high ratio of cells which are unable to form colonies even in a common medium such as TSA was observed. This fact has not been reported previously for common media through recovery after heat shock has been widely studied. After the treatment, staphylococcal protein A, but no enterotoxin, was seen to be released into the extracellular environment.     

Chemotherapeutic treatment for leukemia in a bearded dragon (Pogona vitticeps)

A 4.5-yr-old, captive-bred, male bearded dragon (Pogona vitticeps) presented for lethargy, anorexia, and increased mucoid salivation with upper respiratory clicks. Diagnostics were declined and the bearded dragon was prescribed ceftazidime 20 mg/kg i.m. q 72 hr. The patient presented again 1 wk later with a marked monocytosis, heterophilia, and lymphocytosis, and a clinical diagnosis of chronic monocytic leukemia was made. Chemotherapy with cytosine arabinoside (100 mg/m2 over 48 hr i.v.) was initiated. Forty-four hours into the treatment the dragon became acutely unresponsive and died within 1 hr. Adverse effects as a result of i.v. cytosine arabinoside therapy were not identified despite previous reports suggestive that the drug induces renal failure.     

Proteolytic activity in Hysterothylacium aduncum (Nematoda: Anisakidae), a fish gastrointestinal parasite of worldwide distribution

Proteases have a significant role in the life cycle of parasites and the pathogen-host relationship, being regarded as important virulence factors. In the parasitic nematode Hysterothylacium aduncum proteolytic activity was measured during in vitro development from third larval stage (L3) to mature adult, using DQ red casein as a fluorogenic substrate. Proteolytic activity was detected in all the developmental stages studied and at all pH values within the range employed (2.0-7.5). The assay with specific inhibitors permitted the determination of metalloprotease activity, and, to a lesser extent, that of aspartate- and cysteine-protease. Serine-protease activity was the lowest of those studied. In L3 recently collected from the host fish (L3-0 h), the greatest activity was found at an optimum pH of 4.0 and was mainly inhibited by 1,10-phenathroline (metalloprotease inhibitor). This metalloprotease activity in L3-0 h (infective stage) may be related to the invasion of the host tissues by this larva. In the other developmental stages, the greatest protease activity was found at pH 5.5, although at pH 4.0 a lower activity peak was detected. On the other hand, our data show that the proteolytic activity of the nematode varies according to the presence of pepsin (an aspartic-protease) in the culture medium. Thus, at pH 4.0, activity was greater in the absence of pepsin, with increasing aspartic-protease activity. Together with the detection of aspartic-, cysteine- and metallo-protease (enzymes involved in digestion in invertebrates) in all the developmental stages of the parasite taking place in the digestive tract of the host fish, this allows us to suggest that the pepsin in the culture medium mimics the predigestion conditions in the habitat of the worm within the host and that the activity detected may have, amongst others, a digestive function.     

Seroprevalence of spotted fever group rickettsiae in canines along the United States–Mexico border

Portions of northern Mexico are experiencing a re‐emergence of Rocky Mountain spotted fever (RMSF), a tickborne disease caused by Rickettsia rickettsii, a member of the spotted fever group of rickettsiae (SFGR). Infection with R. rickettsii can result in serious and life‐threatening illness in people and dogs. Canine seroprevalence has been used as a sentinel for human RMSF in previous studies. This study aims to quantify SFGR seroprevalence in canines in three northern Mexican states and identify risk factors associated with seropositivity. A total of 1,136 serum samples and 942 ticks were obtained from dogs participating in government sterilization campaigns and from animal control facilities in 14 Mexican cities in three states. SFGR antibodies were detected using indirect immunofluorescence antibody assays at titre values ≥1/64. Six per cent (69 dogs) showed antibodies to SFGR, with the highest seroprevalence reported in Baja California (12%), Coahuila (4%) and Sonora (4%). Dogs from Baja California had three times higher odds of having SFGR antibodies compared to dogs from Sonora (OR = 3.38, 95% CI, 1.81–6.37). Roughly one quarter (25%) of surveyed dogs were parasitized by ticks (Rhipicephalus sanguineus sensu lato) at the time of sample collection. A portion of collected ticks were tested for rickettsial DNA using polymerase chain reaction. Positive samples were then sequenced, showing evidence of SFGR including R. massiliae, R. parkeri and R. rickettsii. Dogs that spent the majority of time on the street, such as free‐roaming or community‐owned dogs, showed a greater risk of tick infestation, seropositivity, bearing seropositive ticks, and may play a pivotal role in the spread of SFGR among communities. Estimating the seroprevalence of SFGR in the canine population can help public health campaigns target high‐risk communities for interventions to reduce human RMSF cases.     

Medullary plasma pharmacokinetics of vancomycin after intravenous and intraosseous perfusion of the proximal phalanx in horses

OBJECTIVE: To study the pharmacokinetics of vancomycin in plasma obtained from the medullary sinusoids of the proximal phalanx (P1) after intravenous (IV) and intraosseous (IO) regional limb perfusion (RLP). STUDY DESIGN: Experimental study. ANIMALS: Twelve horses. METHODS: IV and IO RLP were performed in 2 groups (n = 6) of horses. Vancomycin hydrochloride (300 mg in 60 mL 0.9% NaCl) was randomly infused in 1 front limb, with the contralateral limb as control (60 mL 0.9% NaCl). A hole was drilled in the lateral cortex of P1, and blood samples from the medullary sinusoids of P1 were collected before infusion, and 15, 30, 45, 65, and 90 minutes after beginning infusion. Samples were centrifuged and plasma vancomycin concentrations determined. Vancomycin concentrations were compared over time and between routes using a 2-way repeated measures ANOVA. Pharmacokinetic variables were compared with a Kolmogorov-Smirnov test. Significance was set at P<.05. RESULTS: No vancomycin-induced clinical side effects such as lameness or swelling were observed. Both techniques, IV and IO, produced high vancomycin concentrations in the sinusoidal plasma of the P1, which remained above the minimum inhibitory concentration value for methicillin-resistant Staphylococcus aureus (MRSA) during the 90 minutes study. Concentrations and pharmacokinetic variables were not significantly different comparing both routes. CONCLUSIONS: IV and IO routes produced similar antimicrobial perfusion of the medullary cavity of P1. CLINICAL RELEVANCE: Either IV or IO routes for perfusion are likely to be equally selected when planning RLP with vancomycin.     

Aeroplysinin-1, a Sponge-Derived Multi-Targeted Bioactive Marine Drug

Organisms lacking external defense mechanisms have developed chemical defense strategies, particularly through the production of secondary metabolites with antibiotic or repellent effects. Secondary metabolites from marine organisms have proven to be an exceptionally rich source of small molecules with pharmacological activities potentially beneficial to human health. (+)-Aeroplysinin-1 is a secondary metabolite isolated from marine sponges with a wide spectrum of bio-activities. (+)-Aeroplysinin-1 has potent antibiotic effects on Gram-positive bacteria and several dinoflagellate microalgae causing toxic blooms. In preclinical studies, (+)-aeroplysinin-1 has been shown to have promising anti-inflammatory, anti-angiogenic and anti-tumor effects. Due to its versatility, (+)-aeroplysinin-1 might have a pharmaceutical interest for the treatment of different pathologies.     

Effect of Temperature on Growth and Survival in Juvenile Opossum Pipefish,Microphis brachyurus: First Observations on the Species in Culture Conditions

In Mexico, natural populations of the opossum pipefish, Microphys brachyurus, are under pressure by anthropogenic alteration of their habitat and unregulated fishing as this species is traded as an aquarium fish, where its survival is low due to inadequate culture practices. The development of culturing techniques has been a successful strategy to protect many exploited species through providing an alternative source to wild exploitation. The aim of this study was to examine the effect on growth, condition, and survival of pipefish cultured for 6 wk at temperatures of 26, 28, and 30 C. Pipefish were fed enriched Artemia nauplii maintaining a ration rate of 14% body weight per day (dry weight Artemia : wet weight fish). The pipefish cultured at 26 and 28 C were longer, heavier, grew faster, and presented better condition than those at 30 C. The survival of pipefish cultured at 26 C was greater (83%) than at 30 C (30%) but not different than at 28 C (50%). This study is the first to report the acceptance of Artemia nauplii by juveniles of the species and that a temperature of 30 C is not suitable for juvenile M. brachyurus culture.