BackgroundAlthough they are important disease vectors mosquito biodiversity in Pakistan is poorly known. Recent epidemics of dengue fever have revealed the need for more detailed understanding of the diversity and distributions of mosquito species in this region. DNA barcoding improves the accuracy of mosquito inventories because morphological differences between many species are subtle, leading to misidentifications.Methodology/Principal FindingsSequence variation in the barcode region of the mitochondrial COI gene was used to identify mosquito species, reveal genetic diversity, and map the distribution of the dengue-vector species in Pakistan. Analysis of 1684 mosquitoes from 491 sites in Punjab and Khyber Pakhtunkhwa during 2010–2013 revealed 32 species with the assemblage dominated by Culex quinquefasciatus (61% of the collection). The genus Aedes (Stegomyia) comprised 15% of the specimens, and was represented by six taxa with the two dengue vector species, Ae. albopictus and Ae. aegypti, dominant and broadly distributed. Anopheles made up another 6% of the catch with An. subpictus dominating. Barcode sequence divergence in conspecific specimens ranged from 0–2.4%, while congeneric species showed from 2.3–17.8% divergence. A global haplotype analysis of disease-vectors showed the presence of multiple haplotypes, although a single haplotype of each dengue-vector species was dominant in most countries. Geographic distribution of Ae. aegypti and Ae. albopictus showed the later species was dominant and found in both rural and urban environments.ConclusionsAs the first DNA-based analysis of mosquitoes in Pakistan, this study has begun the construction of a barcode reference library for the mosquitoes of this region. Levels of genetic diversity varied among species. Because of its capacity to differentiate species, even those with subtle morphological differences, DNA barcoding aids accurate tracking of vector populations.

A key is provided for the 64 recognized species of Podospora. The genus is restricted to species in which the ascospores have an apical germ pore, a basal hyaline cell, and gelatinous appendages. Illustrations are given for 56 species. Detailed descriptions are included for species which are not readily available and the remaining species are briefly described. The following 10 new species found on dung are described and illustrated: P. attoeochaeta from Mexico; P. didyma, Quebec; P. dolichopodalis, Louisiana and Mexico; P. gigantea, Louisiana and Arkansas; P. karachiensis, Pakistan; P. macropodalis, Mexico; P. mexicana, Mexico; P. miniglutinans, Mexico; P. pistilata, Mexico; P. venezuelensis, Venezuela, Mexico, Louisiana, South Pacific. The following new combinations are proposed: P. aloides (Sordaria aloides Fuckel), P. apiculifera (S. apiculifera Speg.), P. argentinensis (S. argentinensis Speg.), P. austro-americana (Hypocopra austro-americana Speg.), P. ellisiana (Pleurage ellisiana Griff.), P. longicollis (Schizothecium longicollis Ames), P. seminuda (Sordaria seminuda Griff.), P. tarvisina (Philocopra tarvisina Sacc.), and P. vesticola (Sphaeria vesticola Berk. & Br.).

The oriental red spider mite, Eutetranychus orientalis (Klein) is a major pest of citrus in many countries including Saudi Arabia (SA). The morphological variations among the different populations of E. orientalis were reported. In the present study, phenotypic variations based on 40 different morphological characteristics were evaluated in 10 E. orientalis populations collected from different hosts and regions of SA. Further, ITS2-rDNA sequences were used to confirm the identity of these phenotypically varying populations. Phenotypic variations in all populations were found in the shape and length of dorsal setae, striation pattern between setae d1 and e1, and leg chaetotaxy. The rDNA sequence analysis of these morphologically varying populations resulted in 10 different ITS2 Saudi haplotypes. The phenotypic and genetic variations were more related to the host plants rather than their geographic distribution. The E. orientalis population collected from Phoenix dactylifera was phenotypically distinct and genetically divergent. The populations collected from citrus species were also more phenotypically and genetically related to each other than to populations collected from non-citrus host plants. The haplotypes recovered from Ziziphus sp., Morus sp., and Azadirecta indica from different regions were grouped in the same sub-clade. Further, the ITS2 haplotypes of Saudi E. orientalis recovered from Citrus reticulata from Riyadh and Al Ula were 100% identical to the ITS2 haplotypes recovered from Citrus sp. from Israel. It is concluded that phenotypic variations exist among different populations of E. orientalis inhabiting different host plants. This species should be identified carefully by considering phenotypic intraspecific variations.

Apodospora is a new coprophilous genus of the Sordariaceae with dark, ostiolate perithecia. The eight-spored ascus has a conspicuous thickened ring in the apex. The asci are mixed with abundant filiform paraphyses. The ascospore has one dark cell and no hyaline cells. They are surrounded by a gelatinous sheath and have a single apical germ pore. The three new species described and illustrated are A. simulans (type of genus), on moose dung, Ontario, Alberta, Idaho, and South Dakota; A. thescelina, on rabbit dung, Wyoming; and A. viridis on rabbit dung, Mexico.

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