Whenever gene expression is being examined, it is essential that a normalization process is carried out to eliminate non-biological variations. The use of reference genes, such as glyceraldehyde-3-phosphate dehydrogenase, actin, and ribosomal protein genes, is the usual method of choice for normalizing gene expression. Although reference genes are used to normalize target gene expression, a major problem is that the stability of these genes differs among tissues, developmental stages, species, and responses to abiotic factors. Therefore, the use and validation of multiple reference genes are required. This review discusses the reasons that why RT-qPCR has become the preferred method for validating results of gene expression profiles, the use of specific and non-specific dyes and the importance of use of primers and probes for qPCR as well as to discuss several statistical algorithms developed to help the validation of potential reference genes. The conflicts arising in the use of classical reference genes in gene normalization and their replacement with novel references are also discussed by citing the high stability and low stability of classical and novel reference genes under various biotic and abiotic experimental conditions by employing various methods applied for the reference genes amplification.

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A new suctorian ciliate, Tokophrya huangmeiensis sp. n. was isolated from the carapace of redclaw crayfish Cherax quadricarinatus, during investigation of fish ciliates diversity in freshwater aquaculture of Hubei, China. The zooids of newly isolated ciliate were found aggregated into colonies on stalks of another ciliate Epistylis. The adults of this newly found species were characterized by an elongated pyramidal shaped and corrugated cell body with two fascicles of tentacles on the apical part of cell body. Stalks were hollow and transparent with longitudinal striations. The single contractile vacuole was located in the apical region and an elongated ρ-shaped macronucleus was situated along the long axis of cell body. Phylogenetic analyses showed that the subclass Suctoria is monophyletic and comprises the groups Exogenina, Evaginogenina and Endogenina, where Exogenina is the earliest branching lineage among these three groups. Tokophrya huangmeiensis sp. n. clustered with the other Tokophrya species and within the Endogenina subgroup as expected.

Small ruminants, especially goats, play an important role in the livestock economy of the world. Parasitism is one of the major problems of the goat population around the world and is responsible for production losses. To control such kinds of losses and improve production, a timely diagnosis of parasitic infection is crucial. The improvement and development of conventional and molecular diagnostic tests help a lot for the early detection of parasitic infections. Strategies to use different control measures like chemotherapy, phytotherapy, pasture management, use of bioactive crops, biological control measures, and development of vaccines also help to improve the health and production of goats.