[{
  "type": "article-journal",
  "title": "MORPHOLOGICAL AND MOLECULAR CONFIRMATION OF THE CHALKBROOD (Ascosphaera apis) INFECTION IN HONEY BEE COLONIES",
  "author": [
    {
      "family": "Muzafar",
      "given": ""
    },
    {
      "family": "Aziz",
      "given": ""
    },
    {
      "family": "Iqbal",
      "given": ""
    },
    {
      "family": "Qayyum",
      "given": ""
    },
    {
      "family": "Alam",
      "given": ""
    },
    {
      "family": "Siddique",
      "given": ""
    },
    {
      "family": "Siddiqui",
      "given": ""
    }
  ],
  "issued": {
    "date-parts": [[2026]]
  },
  "container-title": "Journal of Animal and Plant Sciences",
  "ISSN": "1018-7081",
  "volume": "36",
  "issue": "6",
  "DOI": "https://doi.org/10.36899/JAPS.2026.6.0134",
  "abstract": "<p style=\"text-align: justify;\">Infection by the fungal pathogen&nbsp;<em>Ascosphaera apis</em>&nbsp;causes chalkbrood, a major larval disease that threatens colony health and productivity of honey bee (<em>Apis mellifera&nbsp;</em>L.) globally. This study aimed to determine the prevalence and molecular identity of the pathogen and to access its phylogenetic relationship with internationally reported isolates. In this study,&nbsp;<em>A. apis</em>&nbsp;was identified using morphological and molecular approaches, and the prevalence of chalkbrood infection was assessed in 140 bee colonies. Analyses of chalkbrood mummies revealed that 40 colonies (28.57%) were infected with&nbsp;<em>Ascosphaera</em>&nbsp;spores. Pure isolates cultured on Potato Dextrose Agar (PDA), and PCR-based amplification using ITS gene-specific primers produced a specific 485 bp band, confirming&nbsp;<em>A. apis</em>&nbsp;in the samples. Sequence analysis confirmed the pathogen&rsquo;s identity, and the sequences of&nbsp;<em>A. apis</em>&nbsp;(PQ199405 and PQ282636)<em>&nbsp;</em>were deposited in GenBank. BLAST and phylogenetic analyses of the ITS region sequences showed close similarity (99.12%) with&nbsp;<em>A. apis</em>&nbsp;isolates reported worldwide. This study provides the first molecular confirmation of&nbsp;<em>A. apis&nbsp;</em>in Pakistan and establishes its phylogenetic relationship with global chalkbrood isolates. A baseline dataset generated in this study is essential for future epidemiological studies and supports the development of effective management strategies against chalkbrood disease in regional apiculture.</p>",
  "publisher": "Pakistan Agricultural Scientists Forum",
  "URL": "https://thejaps.org.pk/AbstractView.aspx?mid=2025-JAPS-1178"
}]