Morphological and molecular characterization of a new autochthonous Trichoderma sp. isolate and its biocontrol efficacy against Alternaria sp.

The development of agriculture requires the use of microorganisms in the management of phytopathogens as a way to compensate for the use of chemical pesticides, in order to produce healthy crops. The objective of this study was to characterize a new isolate of Trichoderma sp. based on morphological and molecular features, and its potential ability to control the pathogen Alternaria sp. The antagonistic isolate was isolated from soil samples of potato fields in Guasave Sinaloa, Mexico, whereas the pathogen was collected from infected apple leaves in the orchard "La Escondida" in Guerrero County, Chihuahua, Mexico. For morphological characterization both fungi were grown on solid PDA medium. DNA of Trichoderma sp. was isolated using the CTAB method and PCR analyses were done using ITS1, ITS4 primers resulting in amplified products of 600 bp. These were sequenced, submitted to Genbank (acc. no. MN950427) and used for further phylogenetic analysis through Bayesian inference approach. Five clades were identified and the polytome topography recovered from clade 4 indicates a high genetic similarity with T. asperellum. A BLAST examination of the resulting sequence in GenBank showed 98.11% similarity with T. asperellum. This result together with the morphological and the phylogenetic analyses indicates that the isolate belongs to Trichoderma asperellum Samuels, Lieckfeldt & Nirenberg. Biocontrol tests of this isolate showed inhibition of Alternaria sp. between 50% and 93%. These results are essential for biodiversity research and give some new possibilities for pest management.

Rehabilitation of Marine Turtles and Welfare Improvement by Application of Environmental Enrichment Strategies.

Sea turtles perform various ecological services in several marine environments and are considered architects of the marine landscape. At present, they are endangered species due to anthropogenic threats, pollution and degradation of marine habitats. These impacts make it urgent to increase protection and conservation efforts. Protective actions include the rescue and rehabilitation of injured individuals as a result of their interactions with humans and other threats. Environmental enrichment (EE) is a series of techniques and methods aimed to improve the welfare of animals in captivity and/or under rehabilitation. It uses external stimuli to enhance their psychological and physiological wellbeing to promote natural abilities and behaviors. These may increase the survival chances of rehabilitated animals upon release in the wild. This review presents data of studies where EE has been applied during the rehabilitation processes of different species of sea turtles, and its effect on welfare improvement during captivity/rehabilitation and on survival after release into nature. Technologies such as satellite tags are an important means to determine rehabilitation success and survival of injured individuals from endangered species after release into the wild, as they allow tracking and monitoring of such individuals, and determine their location in areas used by their natural populations for feeding or breeding.

Mini Review: Virus Interference: History, Types and Occurrence in Crustaceans.

Virus interference is a phenomenon in which two viruses interact within a host, affecting the outcome of infection of at least one of such viruses. The effect of this event was first observed in the XVIII century and it was first recorded even before virology was recognized as a distinct science from microbiology. Studies on virus interference were mostly done in the decades between 1930 and 1960 in viruses infecting bacteria and different vertebrates. The systems included in vivo experiments and later, more refined assays were done using tissue and cell cultures. Many viruses involved in interference are pathogenic to humans or to economically important animals. Thus the phenomenon may be relevant to medicine and to animal production due to the possibility to use it as alternative to chemical therapies against virus infections to reduce the severity of disease/mortality caused by a superinfecting virus. Virus interference is defined as the host resistance to a superinfection caused by a pathogenic virus causing obvious signs of disease and/or mortality due to the action of an interfering virus abrogating the replication of the former virus. Different degrees of inhibition of the superinfecting virus can occur. Due to the emergence of novel pathogenic viruses in recent years, virus interference has recently been revisited using different pathogens and hosts, including commercially important farmed aquatic species. Here, some highly pathogenic viruses affecting farmed crustaceans can be affected by interference with other viruses. This review presents data on the history of virus interference in hosts including bacteria and animals, with emphasis on the known cases of virus interference in crustacean hosts. Life Science Identifiers (LSIDs) Escherichia coli [(Migula 1895) Castellani & Chalmers 1919] Aedes albopictus (Skuse 1894) Liocarcinus depurator (Linnaeus 1758): urn:lsid:marinespecies.org:taxname:107387 Penaeus duorarum (Burkenroad 1939): urn:lsid:marinespecies.org:taxname:158334 Carcinus maenas (Linnaeus 1758): urn:lsid:marinespecies.org:taxname:107381 Macrobrachium rosenbergii (De Man 1879): urn:lsid:marinespecies.org:taxname:220137 Penaeus vannamei (Boone 1931): urn:lsid:zoobank.org:pub:C30A0A50-E309-4E24-851D-01CF94D97F23 Penaeus monodon (Fabricius 1798): urn:lsid:zoobank.org:act:3DD50D8B-01C2-48A7-B80D-9D9DD2E6F7AD Penaeus stylirostris (Stimpson 1874): urn:lsid:marinespecies.org:taxname:584982.

Corrigenda: César Marcial Escobedo-Bonilla, José Luis Ibarra Rangel (2014) Susceptibility to an inoculum of infectious hypodermal and haematopoietic necrosis virus (IHHNV) in three batches of whiteleg shrimp Litopenaeus vannamei (Boone, 1931). ZooKeys 457: 355-365. doi: 10.3897/zookeys.457.6715.

[This corrects the article DOI: 10.3897/zookeys.457.6715.].

Susceptibility to an inoculum of infectious hypodermal and haematopoietic necrosis virus (IHHNV) in three batches of whiteleg shrimp Litopenaeusvannamei (Boone, 1931).

The present study evaluated the susceptibility of three different batches of whiteleg shrimp Litopenaeusvannamei from Mexico to an inoculum of infectious hypodermal and haematopoietic necrosis virus (IHHNV). Each of the three shrimp batches came from a different hatchery. Because of their origin, it was possible that the genetic makeup of these batches was different among each other. The three batches tested showed differences in IHHNV susceptibility. Here, susceptibility is defined as the capacity of the host to become infected, and it can be measured by the infectivity titer. Susceptibility to IHHNV was observed in decreasing order in shrimp from batch 1 (hatchery from El Rosario, Sinaloa), batch 3 (hatchery from Nayarit) and batch 2 (hatchery from El Walamo, Sinaloa), respectively. The largest susceptibility difference between batches was 5012 times, and that between early and late juveniles from the same batch was 25 times. These results indicate that within a species, susceptibility to a pathogen such as IHHNV can have large differences. Susceptibility to pathogens is an important trait to consider before performing studies on pathogenesis. It may influence virological parameters such as speed of replication, pathogenicity and virus titer. In order to evaluate the potential use of IHHNV as a natural control agent against white spot syndrome virus (WSSV), it is necessary to know host susceptibility and the kinetics of IHHNV infection. These features can help to determine the conditions in which IHHNV could be used as antagonist in a WSSV infection.

Double-stranded RNA against white spot syndrome virus (WSSV) vp28 or vp26 reduced susceptibility of Litopenaeus vannamei to WSSV, and survivors exhibited decreased susceptibility in subsequent re-infections.

The antiviral effect of vp28 or vp26 double-stranded (ds) RNA upon single or consecutive white spot syndrome virus (WSSV) intramuscular challenges with a high infectious dose was evaluated. The vp28 dsRNA showed the highest protection both in single (LT(50)=145h at 10d and 98h at 20d post treatment [dpt]) or consecutive (LT(50)=765h) WSSV challenges compared to vp26 dsRNA (LT(50)=126h at 10 d and 57h at 20dpt vs. consecutive challenge LT(50)=751h). Single WSSV challenges showed that animals treated with vp28 or vp26 dsRNA gradually lost the antiviral effect as virus challenge occurred at 10dpt (cumulative mortality 63% vs. 80%, respectively) or 20dpt (87% vs. 100%, respectively). In contrast, animals treated with vp28 or vp26 dsRNA and consecutively challenged with WSSV showed and extended lower susceptibility to WSSV. All dead animals were WSSV-positive by one-step PCR, whereas all surviving shrimp from single or continuous challenges were WSSV-negative as determined by reverse transcription (RT)-PCR. In conclusion, shrimp treated with a single administration of vp28 or vp26 dsRNA and consecutively challenged with WSSV showed a stronger and longer antiviral response than shrimp exposed once to WSSV at 10 or 20dpt.

Detection of white spot syndrome virus (WSSV) in the Pacific oyster Crassostrea gigas.

Oysters Crassostrea gigas were placed at water supply canals of three shrimp farms in Guasave, Mexico where WSSV outbreaks occur. Animals were sampled through April-August and September-December to detect WSSV DNA. By using three different PCR protocols, only oysters from a farm undergoing a WSSV outbreak were found WSSV-positive in gills and digestive gland. Two WSSV amplicons were sequenced and they corresponded over 99% to WSSV genome segments. Results showed that oysters can capture WSSV particles suspended in water. Susceptibility of oysters to WSSV infection and their role as a carrier remain to be determined.