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Ameriserv Financial Inc./PA – ‘10-K’ for 12/31/23 – ‘EX-101.DEF’

On:  Wednesday, 3/27/24, at 5:19pm ET   ·   For:  12/31/23   ·   Accession #:  1558370-24-4100   ·   File #:  0-11204

Previous ‘10-K’:  ‘10-K’ on 3/27/23 for 12/31/22   ·   Latest ‘10-K’:  This Filing   ·   10 References:   

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  As Of               Filer                 Filing    For·On·As Docs:Size             Issuer                      Filing Agent

 3/27/24  Ameriserv Financial Inc./PA       10-K       12/31/23  153:33M                                    Toppan Merrill Bridge/FA

Annual Report   —   Form 10-K   —   SEA’34

Filing Table of Contents

Document/Exhibit                   Description                      Pages   Size 

 1: 10-K        Annual Report                                       HTML   7.86M 
 2: EX-4.1      Instrument Defining the Rights of Security Holders  HTML     57K 
 3: EX-21.1     Subsidiaries List                                   HTML     43K 
 4: EX-23.1     Consent of Expert or Counsel                        HTML     40K 
 9: EX-97       Clawback Policy re: Recovery of Erroneously         HTML     61K 
                Awarded Compensation                                             
 5: EX-31.1     Certification -- §302 - SOA'02                      HTML     47K 
 6: EX-31.2     Certification -- §302 - SOA'02                      HTML     47K 
 7: EX-32.1     Certification -- §906 - SOA'02                      HTML     44K 
 8: EX-32.2     Certification -- §906 - SOA'02                      HTML     45K 
15: R1          Document And Entity Information                     HTML    112K 
16: R2          Consolidated Balance Sheets                         HTML    162K 
17: R3          Consolidated Balance Sheets (Parenthetical)         HTML     62K 
18: R4          Consolidated Statements of Operations               HTML    176K 
19: R5          Consolidated Statements of Comprehensive Income     HTML     84K 
                (Loss)                                                           
20: R6          Consolidated Statements of Changes in               HTML     75K 
                Shareholders' Equity                                             
21: R7          Consolidated Statements of Changes in               HTML     50K 
                Shareholders' Equity (Parenthetical)                             
22: R8          Consolidated Statements of Cash Flows               HTML    166K 
23: R9          Summary of Significant Accounting Policies          HTML    157K 
24: R10         Adoption of Accounting Standards                    HTML    137K 
25: R11         Revenue Recognition                                 HTML     76K 
26: R12         Cash and Due From Depository Institutions           HTML     44K 
27: R13         Investment Securities                               HTML    441K 
28: R14         Loans                                               HTML     93K 
29: R15         Allowance for Credit Losses - Loans                 HTML   1.13M 
30: R16         Premises and Equipment                              HTML     61K 
31: R17         Lease Commitments                                   HTML    130K 
32: R18         Deposits                                            HTML     75K 
33: R19         Short-Term Borrowings                               HTML     80K 
34: R20         Advances From Federal Home Loan Bank and            HTML     72K 
                Subordinated Debt                                                
35: R21         Disclosures About Fair Value Measurements           HTML    198K 
36: R22         Disclosures About Fair Value of Financial           HTML    131K 
                Instruments                                                      
37: R23         Income Taxes                                        HTML    126K 
38: R24         Employee Benefit Plans                              HTML    234K 
39: R25         Commitments and Contingent Liabilities              HTML     48K 
40: R26         Stock Compensation Plans                            HTML    101K 
41: R27         Accumulated Other Comprehensive Loss                HTML    205K 
42: R28         Intangible Assets                                   HTML     65K 
43: R29         Derivative Hedging Instruments                      HTML    118K 
44: R30         Segment Results                                     HTML    156K 
45: R31         Regulatory Capital                                  HTML    158K 
46: R32         Parent Company Financial Information                HTML    182K 
47: R33         Selected Quarterly Consolidated Financial Data      HTML    138K 
                (Unaudited)                                                      
48: R34         Summary of Significant Accounting Policies          HTML    220K 
                (Policies)                                                       
49: R35         Summary of Significant Accounting Policies          HTML     87K 
                (Tables)                                                         
50: R36         Adoption of Accounting Standards (Tables)           HTML    123K 
51: R37         Revenue Recognition (Tables)                        HTML     67K 
52: R38         Investment Securities (Tables)                      HTML    441K 
53: R39         Loans (Tables)                                      HTML     91K 
54: R40         Allowance for Credit Losses - Loans (Tables)        HTML   1.12M 
55: R41         Premises and Equipment (Tables)                     HTML     60K 
56: R42         Lease Commitments (Tables)                          HTML    132K 
57: R43         Deposits (Tables)                                   HTML     74K 
58: R44         Short-Term Borrowings (Tables)                      HTML     80K 
59: R45         Advances From Federal Home Loan Bank and            HTML     68K 
                Subordinated Debt (Tables)                                       
60: R46         Disclosures About Fair Value Measurements (Tables)  HTML    196K 
61: R47         Disclosures About Fair Value of Financial           HTML    129K 
                Instruments (Tables)                                             
62: R48         Income Taxes (Tables)                               HTML    127K 
63: R49         Employee Benefit Plans (Tables)                     HTML    237K 
64: R50         Stock Compensation Plans (Tables)                   HTML     99K 
65: R51         Accumulated Other Comprehensive Loss (Tables)       HTML    207K 
66: R52         Intangible Assets (Tables)                          HTML     65K 
67: R53         Derivative Hedging Instruments (Tables)             HTML    110K 
68: R54         Segment Results (Tables)                            HTML    150K 
69: R55         Regulatory Capital (Tables)                         HTML    154K 
70: R56         Parent Company Financial Information (Tables)       HTML    184K 
71: R57         Selected Quarterly Consolidated Financial Data      HTML    138K 
                (Unaudited) (Tables)                                             
72: R58         SUMMARY OF SIGNIFICANT ACCOUNTING POLICIES -        HTML    163K 
                Additional information (Details)                                 
73: R59         SUMMARY OF SIGNIFICANT ACCOUNTING POLICIES -        HTML     79K 
                Computation of diluted earnings per common share                 
                (Details)                                                        
74: R60         SUMMARY OF SIGNIFICANT ACCOUNTING POLICIES -        HTML     67K 
                Non-cash assets acquired and Liabilities Assumed                 
                (Details)                                                        
75: R61         Adoption of Accounting Standards (Details)          HTML     90K 
76: R62         Adoption of Accounting Standards - Current          HTML    167K 
                Expected Credit Loss Model (Details)                             
77: R63         Revenue Recognition (Details)                       HTML     57K 
78: R64         Revenue Recognition - Additional information        HTML     44K 
                (Details)                                                        
79: R65         Cash and Due From Depository Institutions           HTML     46K 
                (Details)                                                        
80: R66         INVESTMENT SECURITIES - Cost basis and fair values  HTML     91K 
                of investment securities (Details)                               
81: R67         INVESTMENT SECURITIES - Total investment            HTML    132K 
                securities (Details)                                             
82: R68         INVESTMENT SECURITIES - Information concerning      HTML     71K 
                investments with unrealized losses (Details)                     
83: R69         INVESTMENT SECURITIES - Allowance for credit        HTML     63K 
                losses on held to maturity debt securities by                    
                major security type (Details)                                    
84: R70         INVESTMENT SECURITIES - Amortized cost of held to   HTML     67K 
                maturity debt securities (Details)                               
85: R71         INVESTMENT SECURITIES - Additional information      HTML    197K 
                (Details)                                                        
86: R72         LOANS - Loan Portfolio (Details)                    HTML     90K 
87: R73         LOANS - Additional information (Details)            HTML     49K 
88: R74         LOANS - Summary of loan activity with related       HTML     48K 
                parties (Details)                                                
89: R75         ALLOWANCE FOR CREDIT LOSSES - LOANS - Allowance     HTML    133K 
                for credit losses by loan portfolio segment                      
                (Details)                                                        
90: R76         ALLOWANCE FOR CREDIT LOSSES - LOANS - Loan          HTML    128K 
                portfolio and allowance for credit losses                        
                (Details)                                                        
91: R77         ALLOWANCE FOR CREDIT LOSSES - LOANS - Amortized     HTML     78K 
                cost basis of collateral-dependent loans (Details)               
92: R78         ALLOWANCE FOR CREDIT LOSSES - LOANS -               HTML     94K 
                Non-Performing Assets (Details)                                  
93: R79         ALLOWANCE FOR CREDIT LOSSES - LOANS - Commercial    HTML    238K 
                and commercial real estate loan portfolios                       
                (Details)                                                        
94: R80         ALLOWANCE FOR CREDIT LOSSES - LOANS - Residential   HTML    139K 
                and consumer portfolio (Details)                                 
95: R81         ALLOWANCE FOR CREDIT LOSSES - LOANS - Aging         HTML    155K 
                categories of performing loans and non-accrual                   
                loans (Details)                                                  
96: R82         ALLOWANCE FOR CREDIT LOSSES - LOANS - Loan          HTML     80K 
                modifications to borrowers experiencing financial                
                difficulty (Details)                                             
97: R83         ALLOWANCE FOR CREDIT LOSSES - LOANS - Loan          HTML     53K 
                modified as a TDR (Details)                                      
98: R84         ALLOWANCE FOR CREDIT LOSSES - LOANS - Additional    HTML    167K 
                information (Details)                                            
99: R85         Premises and Equipment (Details)                    HTML     57K 
100: R86         Premises and Equipment - Additional information     HTML     48K  
                (Details)                                                        
101: R87         LEASE COMMITMENTS - Lease Cost (Details)            HTML     50K  
102: R88         LEASE COMMITMENTS - Leases outstanding (Details)    HTML     50K  
103: R89         LEASE COMMITMENTS - Operating and financing leases  HTML     80K  
                - (Details)                                                      
104: R90         LEASE COMMITMENTS - Additional information          HTML     51K  
                (Details)                                                        
105: R91         Deposits (Details)                                  HTML     53K  
106: R92         Deposits - Maturity of time deposits (Details)      HTML     55K  
107: R93         DEPOSITS - Additional information (Details)         HTML     44K  
108: R94         Short-Term Borrowings (Details)                     HTML     57K  
109: R95         Advances From Federal Home Loan Bank and            HTML     72K  
                Subordinated Debt (Details)                                      
110: R96         Advances From Federal Home Loan Bank and            HTML     75K  
                Subordinated Debt (Details)                                      
111: R97         DISCLOSURES ABOUT FAIR VALUE MEASUREMENTS -         HTML    103K  
                Schedule of assets and liability measured and                    
                recorded on the Consolidated Balance Sheets                      
                (Details)                                                        
112: R98         DISCLOSURES ABOUT FAIR VALUE MEASUREMENTS -         HTML     96K  
                Schedule of assets measured and recorded at fair                 
                value on a non-recurring basis (Details)                         
113: R99         DISCLOSURES ABOUT FAIR VALUE OF FINANCIAL           HTML     74K  
                INSTRUMENTS - Schedule of estimated fair value and               
                recorded carrying value (Details)                                
114: R100        Income Taxes (Details)                              HTML     52K  
115: R101        Income Taxes - Reconciliation (Details)             HTML     67K  
116: R102        INCOME TAXES - Deferred tax assets and liabilities  HTML     79K  
                (Details)                                                        
117: R103        EMPLOYEE BENEFIT PLANS - Changes in obligations     HTML     78K  
                and assets (Details)                                             
118: R104        EMPLOYEE BENEFIT PLANS - Amounts not recognized as  HTML     47K  
                components of net periodic pension cost (Details)                
119: R105        EMPLOYEE BENEFIT PLANS - Accumulated benefit        HTML     44K  
                obligation (Details)                                             
120: R106        EMPLOYEE BENEFIT PLANS - Weighted-average           HTML     53K  
                assumptions used to determine benefit obligations                
                (Details)                                                        
121: R107        EMPLOYEE BENEFIT PLANS - Component of net periodic  HTML     60K  
                benefit cost (Details)                                           
122: R108        EMPLOYEE BENEFIT PLANS - Changes in plan assets     HTML     52K  
                and benefit obligations (Details)                                
123: R109        EMPLOYEE BENEFIT PLANS - Weighted-average           HTML     58K  
                assumptions used to determine net periodic benefit               
                cost (Details)                                                   
124: R110        EMPLOYEE BENEFIT PLANS - Asset allocation           HTML     52K  
                (Details)                                                        
125: R111        EMPLOYEE BENEFIT PLANS - Segregation of assets by   HTML     57K  
                the level of valuations inputs (Details)                         
126: R112        EMPLOYEE BENEFIT PLANS - Additional information     HTML    122K  
                (Details)                                                        
127: R113        EMPLOYEE BENEFIT PLANS - Estimated future benefit   HTML     54K  
                payments (Details)                                               
128: R114        Commitments and Contingent Liabilities (Details)    HTML     61K  
129: R115        Stock Compensation Plans (Details)                  HTML     71K  
130: R116        Stock Compensation Plans - Pricing model            HTML     55K  
                assumption ranges (Details)                                      
131: R117        STOCK COMPENSATION PLANS - Additional information   HTML     86K  
                (Details)                                                        
132: R118        Accumulated Other Comprehensive Loss (Details)      HTML     68K  
133: R119        Accumulated Other Comprehensive Loss -              HTML     88K  
                Reclassification of component (Details)                          
134: R120        Intangible Assets (Details)                         HTML     63K  
135: R121        Intangible Assets - Amortization of intangible      HTML     49K  
                (Details)                                                        
136: R122        INTANGIBLE ASSETS - Future Amortization (Details)   HTML     59K  
137: R123        Derivative Hedging Instruments (Details)            HTML     59K  
138: R124        Derivative Hedging Instruments - Additional         HTML     57K  
                Information (Details)                                            
139: R125        DERIVATIVE HEDGING INSTRUMENTS - Cash Flow Hedge    HTML     53K  
                (Details)                                                        
140: R126        Segment Results (Details)                           HTML    106K  
141: R127        Regulatory Capital (Details)                        HTML     79K  
142: R128        Parent Company Financial Information - Balance      HTML    103K  
                Sheets (Details)                                                 
143: R129        Parent Company Financial Information - Statements   HTML    115K  
                of Operations (Details)                                          
144: R130        Parent Company Financial Information - Statements   HTML    121K  
                of Cash Flows (Details)                                          
145: R131        PARENT COMPANY FINANCIAL INFORMATION - Additional   HTML     42K  
                information (Details)                                            
146: R132        Selected Quarterly Consolidated Financial Data      HTML     99K  
                (Unaudited) (Details)                                            
147: R133        Pay vs Performance Disclosure                       HTML     54K  
148: R134        Insider Trading Arrangements                        HTML     47K  
150: XML         IDEA XML File -- Filing Summary                      XML    293K  
153: XML         XBRL Instance -- asrv-20231231x10k_htm               XML  10.34M  
149: EXCEL       IDEA Workbook of Financial Report Info              XLSX    381K  
11: EX-101.CAL  XBRL Calculations -- asrv-20231231_cal               XML    359K 
12: EX-101.DEF  XBRL Definitions -- asrv-20231231_def                XML   1.69M 
13: EX-101.LAB  XBRL Labels -- asrv-20231231_lab                     XML   2.40M 
14: EX-101.PRE  XBRL Presentations -- asrv-20231231_pre              XML   2.41M 
10: EX-101.SCH  XBRL Schema -- asrv-20231231                         XSD    326K 
151: JSON        XBRL Instance as JSON Data -- MetaLinks              931±  1.47M  
152: ZIP         XBRL Zipped Folder -- 0001558370-24-004100-xbrl      Zip   1.20M  


‘EX-101.DEF’   —   XBRL Definitions — asrv-20231231_def


This Exhibit is an XBRL File.


                                                                                                                                                                                
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<!-- XBRL document created with Toppan Merrill Bridge 10.5.0.8 -->
<!-- Based on XBRL 2.1 -->
<!-- Created on: 3/27/2024 8:05:40 PM -->
<!-- Modified on: 3/27/2024 8:05:41 PM -->
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<link:roleRef xlink:type="simple" xlink:href="asrv-20231231.xsd#DisclosureAdoptionOfAccountingStandardsDetails" roleURI="http://www.ameriserv.com/role/DisclosureAdoptionOfAccountingStandardsDetails"/>
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<link:roleRef xlink:type="simple" xlink:href="asrv-20231231.xsd#DisclosureRevenueRecognitionDetails" roleURI="http://www.ameriserv.com/role/DisclosureRevenueRecognitionDetails"/>
<link:roleRef xlink:type="simple" xlink:href="asrv-20231231.xsd#DisclosureRevenueRecognitionAdditionalInformationDetails" roleURI="http://www.ameriserv.com/role/DisclosureRevenueRecognitionAdditionalInformationDetails"/>
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2 Subsequent Filings that Reference this Filing

  As Of               Filer                 Filing    For·On·As Docs:Size             Issuer                      Filing Agent

 4/22/24  Ameriserv Financial Inc./PA       DEFA14A                1:145K                                   Toppan Merrill/FA
 4/22/24  Ameriserv Financial Inc./PA       8-K:1,8,9   4/18/24   12:343K                                   Toppan Merrill/FA


8 Previous Filings that this Filing References

  As Of               Filer                 Filing    For·On·As Docs:Size             Issuer                      Filing Agent

 8/29/23  Ameriserv Financial Inc./PA       8-K:5,9     8/28/23   11:360K                                   Toppan Merrill/FA
 3/23/21  Ameriserv Financial Inc./PA       DEF 14A     4/27/21    1:1.2M                                   Toppan Merrill/FA
 4/06/20  Ameriserv Financial Inc./PA       8-K:5,8,9   4/06/20    1:98K
 2/24/16  Ameriserv Financial Inc./PA       8-K:5,9     2/19/16    1:50K
 4/28/15  Ameriserv Financial Inc./PA       8-K:5,9     4/28/15    2:117K
10/21/14  Ameriserv Financial Inc./PA       8-K:5      10/21/14    1:91K
 9/16/11  Ameriserv Financial Inc./PA       S-8         9/16/11    5:185K
 3/21/11  Ameriserv Financial Inc./PA       DEF 14A     4/26/11    1:737K                                   Donnelley … Solutions/FA
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